The document summarizes a study on the prevalence of canine parvovirus (CPV) in domestic dogs living around Serengeti National Park in Tanzania. Blood samples were collected from 77 asymptomatic domestic dogs and tested for CPV using PCR. The results found that 10.4% of samples were positive for CPV, with 6.5% positive for the CPV-2a strain and 3.9% for CPV-2b. This suggests that domestic dogs can act as reservoirs for CPV transmission to other dogs and wildlife in the area.
Identification of SNP markers for resistance to Salmonella and IBDV in indige...ILRI
Poster prepared by Psifidi, G. Banos, O. Matika, Tadelle Dessie, R. Christley, P. Wigley, J.M. Bettridge, O. Hanotte, Takele Taye Desta and P. Kaiser for the Annual Meeting of the Society of Veterinary Epidemiology and Preventive Medicine, Madrid, Spain, 20-22 March 2013.
Variation analysis of Swine influenza virus (SIV) H1N1 sequences in experimen...Álvaro L. Valiñas
Swine influenza is a highly contagious and widely distributed disease that generates important economic losses in the pig industry. Nowadays, one of the most extended strategy used to control Swine influenza viruses (SIVs) is the trivalent vaccine application, which formulation contains the most frequently circulating SIV subtypes H1N1, H1N2 and H3N2. These vaccines do not provide sterilizing immunity against the virus, potentially favoring viral evolutionary dynamics. To better understand the main mechanisms that shape viral evolution, in this work, the SIV intra-host diversity was analyzed in samples collected from both, vaccinated and non-vaccinated animals challenged with H1N1 influenza A virus. In the present study 276 single nucleotide variants were found within 28 whole SIV genomes obtained by next generation sequencing. Differences in nucleotide variants between groups were established and the impact of each substitution found was hypothesized according to previous literature. Substitutions were allocated along all influenza genetic segments, while the most relevant non-synonymous substitutions were allocated in the NS1 protein on samples collected only from vaccinated animals. These substitutions could affect both, mRNA viral translation and pathogenesis. Moreover, new viral variants were found in both vaccinated and non-vaccinated pigs, showing relevant substitutions in the HA, NA and NP proteins that may be contributing to evasion of host immune system, virulence and host adaptation. Overall, results of the present study suggest that SIV is continuously evolving despite vaccine application, therefore new substitutions may increase viral fitness under field conditions.
Variation analysis of Swine influenza virus (SIV) H1N1 sequences in experimen...Álvaro L. Valiñas
Swine influenza is a highly contagious and widely distributed disease that generates important economic losses in the pig industry. Nowadays, one of the most extended strategy used to control Swine influenza viruses (SIVs) is the trivalent vaccine application, which formulation contains the most frequently circulating SIV subtypes H1N1, H1N2 and H3N2. These vaccines do not provide sterilizing immunity against the virus, potentially favoring viral evolutionary dynamics. To better understand the main mechanisms that shape viral evolution, in this work, the SIV intra-host diversity was analyzed in samples collected from both, vaccinated and non-vaccinated animals challenged with H1N1 influenza A virus. In the present study 276 single nucleotide variants were found within 28 whole SIV genomes obtained by next generation sequencing. Differences in nucleotide variants between groups were established and the impact of each substitution found was hypothesized according to previous literature. Substitutions were allocated along all influenza genetic segments, while the most relevant non-synonymous substitutions were allocated in the NS1 protein on samples collected only from vaccinated animals. These substitutions could affect both, mRNA viral translation and pathogenesis. Moreover, new viral variants were found in both vaccinated and non-vaccinated pigs, showing relevant substitutions in the HA, NA and NP proteins that may be contributing to evasion of host immune system, virulence and host adaptation. Overall, results of the present study suggest that SIV is continuously evolving despite vaccine application, therefore new substitutions may increase viral fitness under field conditions.
A presentation as a webinar for the Winn Feline Foundation that focuses on recent findings related to the signatures of selection in the domestic cat genome
Sero-evidence of zoonotic viruses in rodents and humans in Kibera informal se...ILRI
Poster prepared Joseph Ogola, Hussein Alburkat, Moses Masika, Essi Korhonen, Ruut Uusitalo, Philip Nyaga, Omu Anzala, Olli Vapalahti, Tarja Sironen and Kristian M. Forbes for the Kenya One Health Online Conference, 6-8 December 2021
Identification of SNP markers for resistance to Salmonella and IBDV in indige...ILRI
Poster prepared by Psifidi, G. Banos, O. Matika, Tadelle Dessie, R. Christley, P. Wigley, J.M. Bettridge, O. Hanotte, Takele Taye Desta and P. Kaiser for the Annual Meeting of the Society of Veterinary Epidemiology and Preventive Medicine, Madrid, Spain, 20-22 March 2013.
Variation analysis of Swine influenza virus (SIV) H1N1 sequences in experimen...Álvaro L. Valiñas
Swine influenza is a highly contagious and widely distributed disease that generates important economic losses in the pig industry. Nowadays, one of the most extended strategy used to control Swine influenza viruses (SIVs) is the trivalent vaccine application, which formulation contains the most frequently circulating SIV subtypes H1N1, H1N2 and H3N2. These vaccines do not provide sterilizing immunity against the virus, potentially favoring viral evolutionary dynamics. To better understand the main mechanisms that shape viral evolution, in this work, the SIV intra-host diversity was analyzed in samples collected from both, vaccinated and non-vaccinated animals challenged with H1N1 influenza A virus. In the present study 276 single nucleotide variants were found within 28 whole SIV genomes obtained by next generation sequencing. Differences in nucleotide variants between groups were established and the impact of each substitution found was hypothesized according to previous literature. Substitutions were allocated along all influenza genetic segments, while the most relevant non-synonymous substitutions were allocated in the NS1 protein on samples collected only from vaccinated animals. These substitutions could affect both, mRNA viral translation and pathogenesis. Moreover, new viral variants were found in both vaccinated and non-vaccinated pigs, showing relevant substitutions in the HA, NA and NP proteins that may be contributing to evasion of host immune system, virulence and host adaptation. Overall, results of the present study suggest that SIV is continuously evolving despite vaccine application, therefore new substitutions may increase viral fitness under field conditions.
Variation analysis of Swine influenza virus (SIV) H1N1 sequences in experimen...Álvaro L. Valiñas
Swine influenza is a highly contagious and widely distributed disease that generates important economic losses in the pig industry. Nowadays, one of the most extended strategy used to control Swine influenza viruses (SIVs) is the trivalent vaccine application, which formulation contains the most frequently circulating SIV subtypes H1N1, H1N2 and H3N2. These vaccines do not provide sterilizing immunity against the virus, potentially favoring viral evolutionary dynamics. To better understand the main mechanisms that shape viral evolution, in this work, the SIV intra-host diversity was analyzed in samples collected from both, vaccinated and non-vaccinated animals challenged with H1N1 influenza A virus. In the present study 276 single nucleotide variants were found within 28 whole SIV genomes obtained by next generation sequencing. Differences in nucleotide variants between groups were established and the impact of each substitution found was hypothesized according to previous literature. Substitutions were allocated along all influenza genetic segments, while the most relevant non-synonymous substitutions were allocated in the NS1 protein on samples collected only from vaccinated animals. These substitutions could affect both, mRNA viral translation and pathogenesis. Moreover, new viral variants were found in both vaccinated and non-vaccinated pigs, showing relevant substitutions in the HA, NA and NP proteins that may be contributing to evasion of host immune system, virulence and host adaptation. Overall, results of the present study suggest that SIV is continuously evolving despite vaccine application, therefore new substitutions may increase viral fitness under field conditions.
A presentation as a webinar for the Winn Feline Foundation that focuses on recent findings related to the signatures of selection in the domestic cat genome
Sero-evidence of zoonotic viruses in rodents and humans in Kibera informal se...ILRI
Poster prepared Joseph Ogola, Hussein Alburkat, Moses Masika, Essi Korhonen, Ruut Uusitalo, Philip Nyaga, Omu Anzala, Olli Vapalahti, Tarja Sironen and Kristian M. Forbes for the Kenya One Health Online Conference, 6-8 December 2021
Reseach on H9N2: evidence that link outbreaks in Eurasia, China, South Korea,...Harm Kiezebrink
In this study, scientists from the U.S. Geological Survey and U.S. Fish and Wildlife Service harnessed a new type of DNA technology to investigate avian influenza viruses in Alaska. Using a “next generation” sequencing approach, which identifies gene sequences of interest more rapidly and more completely than by traditional techniques, scientists identified low pathogenic avian influenza viruses in Alaska that are nearly identical to viruses found in China and South Korea.
The viruses were found in an area of western Alaska that is known to be a hot spot for both American and Eurasian forms of avian influenza.
“Our past research in western Alaska has shown that 70 percent of avian influenza viruses isolated in this area were found to contain genetic material from Eurasia, providing evidence for high levels of intercontinental viral exchange,” said Andy Ramey, a scientist with the USGS Alaska Science Center and lead author of the study. “This is because Asian and North American migratory flyways overlap in western Alaska.”
The new study, led by the USGS, found low pathogenic H9N2 viruses in an Emperor Goose and a Northern Pintail. Both of the H9N2 viruses were nearly identical genetically to viruses found in wild bird samples from Lake Dongting, China and Cheon-su Bay, South Korea.
“These H9N2 viruses are low pathogenic and not known to infect humans, but similar viruses have been implicated in disease outbreaks in domestic poultry in Asia,” said Ramey.
There is no commercial poultry production in western Alaska and highly similar H9N2 virus strains have not been reported in poultry in East Asia or North America, so it is unlikely that agricultural imports influenced this result.
The finding provides evidence for intercontinental movement of intact avian influenza viruses by migratory birds. The USGS recently released a publication about the detection of a novel highly pathogenic H5N8 virus in the U.S. that is highly similar to the Eurasian H5N8 viruses. This suggests that the novel re-assortment may be adapted to certain waterfowl species, enabling it to survive long migrations. That virus, and associated strains, have now spread from early detections in wild and domestic birds in Pacific states to poultry outbreaks in Minnesota, Missouri and Arkansas.
“The frequency of inter-hemispheric dispersal events of avian influenza viruses by migratory birds may be higher than previously recognized,” said Ramey.
While some of the samples for the project came from bird fecal samples collected from beaches at Izembek National Wildlife Refuge, most of the samples came from sport hunters.
“For the past several years, we’ve worked closely with sport hunters in the fall to obtain swab samples from birds and that has really informed our understanding of wildlife disease in this area,” said Bruce Casler, formerly a biologist with the USFWS Izembek National Wildlife Refuge and a co-author of the study. Non
Spatio temporal dynamics of global H5N1 outbreaks match bird migration patternsHarm Kiezebrink
The global spread of highly pathogenic avian influenza H5N1 in poultry, wild birds and humans, poses a significant pandemic threat and a serious public health risk.
An efficient surveillance and disease control system relies on the understanding of the dispersion patterns and spreading mechanisms of the virus. A space-time cluster analysis of H5N1 outbreaks was used to identify spatio-temporal patterns at a global scale and over an extended period of time.
Potential mechanisms explaining the spread of the H5N1 virus, and the role of wild birds, were analyzed. Between December 2003 and December 2006, three global epidemic phases of H5N1 influenza were identified.
These H5N1 outbreaks showed a clear seasonal pattern, with a high density of outbreaks in winter and early spring (i.e., October to March). In phase I and II only the East Asia Australian flyway was affected. During phase III, the H5N1 viruses started to appear in four other flyways: the Central Asian flyway, the Black Sea Mediterranean flyway, the East Atlantic flyway and the East Africa West Asian flyway.
Six disease cluster patterns along these flyways were found to be associated with the seasonal migration of wild birds. The spread of the H5N1 virus, as demonstrated by the space-time clusters, was associated with the patterns of migration of wild birds. Wild birds may therefore play an important role in the spread of H5N1 over long distances.
Disease clusters were also detected at sites where wild birds are known to overwinter and at times when migratory birds were present. This leads to the suggestion that wild birds may also be involved in spreading the H5N1 virus over short distances.
Ebola virus surveillance in pigs presenting for slaughter in UgandaILRI
Poster by Christine Atherstone, Silvia Alonso, Delia Grace, Michael Ward, Navneet Dhand and Siobhan Mor presented at the 4th International One Health Congress and 6th Biennial Congress of the International Association for Ecology and Health (One Health EcoHealth 2016), Melbourne, Australia, 3–7 December 2016.
— Herpesviruses that infect fishes belong to the Herpesvirales order and Alloherpesvirus family. In these species, the different types of herpesvirus can cause tumors, adenocarcinoma and skin lesions. This study aims detect to presence of herpesvirus in fishes from commercial, recreation or experimental creations of the States of São Paulo and Minas Gerais, Brazil. Organ fragments and lesions of 53 fish species coming of mortality cases were forwarded at Biological Institute for examination by transmission electron microscopy by research of etiological agent. By transmission electron microscopy through negative staining technique, were observed herpes virus-like particles in 46 fishes and through embedding resin technique, in ultrathin sections were visualized herpes virus immature particles, measuring 90-110nm in diameter, located in the nuclei and complete particles measuring 160nm. In the histopathology technique, lesions associated with the virus as corpuscles inclusion, papillomas, and dermal lesions and in the gills were observed in 27 fishes. The evaluated techniques of TEM and the histopathology were effective for the rapid detection of herpesvirus in the examined samples.
Seroprevalence of Peste des Petits Ruminants in Traditional Goats Reared in N...Premier Publishers
This study aims at determining the prevalence of Peste des Petits Ruminants (PPR) in goats reared traditionally in northern Côte d’Ivoire villages. For that, serum samples collected from 171 goats randomly selected from five localities in the Departement of Korhogo and tested the presence of anti-PPRV antibodies by a Competitive Enzyme-linked Immunosorbent Assay (c-ELISA). Overall, seroprevalence of PPR in the area was 36.26% (62/171). All the localities sampled had at least one PPR-positive animal. Age and sex of the animals were not significantly (p>0.05) associated with the infection; however, localities of sampled animals, showed significant (p<0.05) association with PPR virus-infection in goats. It is then concluded that there is high seroprevalence of PPR in traditional raised goats in northern Côte d’Ivoire. Therefore, vaccination campaigns against PPR are advocated to prevent the transmission and spread of PPR in the area.
Sero-epidemiological investigation of foot and mouth disease in cattle at the...ILRI
Poster by Daniel Nthiwa, Silvia Alonso, David Odongo, Eucharia Kenya and Bernard Bett presented at the 15th International Symposium of Veterinary Epidemiology and Ecology, Chiang Mai, Thailand, 12–16 November 2018.
Reseach on H9N2: evidence that link outbreaks in Eurasia, China, South Korea,...Harm Kiezebrink
In this study, scientists from the U.S. Geological Survey and U.S. Fish and Wildlife Service harnessed a new type of DNA technology to investigate avian influenza viruses in Alaska. Using a “next generation” sequencing approach, which identifies gene sequences of interest more rapidly and more completely than by traditional techniques, scientists identified low pathogenic avian influenza viruses in Alaska that are nearly identical to viruses found in China and South Korea.
The viruses were found in an area of western Alaska that is known to be a hot spot for both American and Eurasian forms of avian influenza.
“Our past research in western Alaska has shown that 70 percent of avian influenza viruses isolated in this area were found to contain genetic material from Eurasia, providing evidence for high levels of intercontinental viral exchange,” said Andy Ramey, a scientist with the USGS Alaska Science Center and lead author of the study. “This is because Asian and North American migratory flyways overlap in western Alaska.”
The new study, led by the USGS, found low pathogenic H9N2 viruses in an Emperor Goose and a Northern Pintail. Both of the H9N2 viruses were nearly identical genetically to viruses found in wild bird samples from Lake Dongting, China and Cheon-su Bay, South Korea.
“These H9N2 viruses are low pathogenic and not known to infect humans, but similar viruses have been implicated in disease outbreaks in domestic poultry in Asia,” said Ramey.
There is no commercial poultry production in western Alaska and highly similar H9N2 virus strains have not been reported in poultry in East Asia or North America, so it is unlikely that agricultural imports influenced this result.
The finding provides evidence for intercontinental movement of intact avian influenza viruses by migratory birds. The USGS recently released a publication about the detection of a novel highly pathogenic H5N8 virus in the U.S. that is highly similar to the Eurasian H5N8 viruses. This suggests that the novel re-assortment may be adapted to certain waterfowl species, enabling it to survive long migrations. That virus, and associated strains, have now spread from early detections in wild and domestic birds in Pacific states to poultry outbreaks in Minnesota, Missouri and Arkansas.
“The frequency of inter-hemispheric dispersal events of avian influenza viruses by migratory birds may be higher than previously recognized,” said Ramey.
While some of the samples for the project came from bird fecal samples collected from beaches at Izembek National Wildlife Refuge, most of the samples came from sport hunters.
“For the past several years, we’ve worked closely with sport hunters in the fall to obtain swab samples from birds and that has really informed our understanding of wildlife disease in this area,” said Bruce Casler, formerly a biologist with the USFWS Izembek National Wildlife Refuge and a co-author of the study. Non
Spatio temporal dynamics of global H5N1 outbreaks match bird migration patternsHarm Kiezebrink
The global spread of highly pathogenic avian influenza H5N1 in poultry, wild birds and humans, poses a significant pandemic threat and a serious public health risk.
An efficient surveillance and disease control system relies on the understanding of the dispersion patterns and spreading mechanisms of the virus. A space-time cluster analysis of H5N1 outbreaks was used to identify spatio-temporal patterns at a global scale and over an extended period of time.
Potential mechanisms explaining the spread of the H5N1 virus, and the role of wild birds, were analyzed. Between December 2003 and December 2006, three global epidemic phases of H5N1 influenza were identified.
These H5N1 outbreaks showed a clear seasonal pattern, with a high density of outbreaks in winter and early spring (i.e., October to March). In phase I and II only the East Asia Australian flyway was affected. During phase III, the H5N1 viruses started to appear in four other flyways: the Central Asian flyway, the Black Sea Mediterranean flyway, the East Atlantic flyway and the East Africa West Asian flyway.
Six disease cluster patterns along these flyways were found to be associated with the seasonal migration of wild birds. The spread of the H5N1 virus, as demonstrated by the space-time clusters, was associated with the patterns of migration of wild birds. Wild birds may therefore play an important role in the spread of H5N1 over long distances.
Disease clusters were also detected at sites where wild birds are known to overwinter and at times when migratory birds were present. This leads to the suggestion that wild birds may also be involved in spreading the H5N1 virus over short distances.
Ebola virus surveillance in pigs presenting for slaughter in UgandaILRI
Poster by Christine Atherstone, Silvia Alonso, Delia Grace, Michael Ward, Navneet Dhand and Siobhan Mor presented at the 4th International One Health Congress and 6th Biennial Congress of the International Association for Ecology and Health (One Health EcoHealth 2016), Melbourne, Australia, 3–7 December 2016.
— Herpesviruses that infect fishes belong to the Herpesvirales order and Alloherpesvirus family. In these species, the different types of herpesvirus can cause tumors, adenocarcinoma and skin lesions. This study aims detect to presence of herpesvirus in fishes from commercial, recreation or experimental creations of the States of São Paulo and Minas Gerais, Brazil. Organ fragments and lesions of 53 fish species coming of mortality cases were forwarded at Biological Institute for examination by transmission electron microscopy by research of etiological agent. By transmission electron microscopy through negative staining technique, were observed herpes virus-like particles in 46 fishes and through embedding resin technique, in ultrathin sections were visualized herpes virus immature particles, measuring 90-110nm in diameter, located in the nuclei and complete particles measuring 160nm. In the histopathology technique, lesions associated with the virus as corpuscles inclusion, papillomas, and dermal lesions and in the gills were observed in 27 fishes. The evaluated techniques of TEM and the histopathology were effective for the rapid detection of herpesvirus in the examined samples.
Seroprevalence of Peste des Petits Ruminants in Traditional Goats Reared in N...Premier Publishers
This study aims at determining the prevalence of Peste des Petits Ruminants (PPR) in goats reared traditionally in northern Côte d’Ivoire villages. For that, serum samples collected from 171 goats randomly selected from five localities in the Departement of Korhogo and tested the presence of anti-PPRV antibodies by a Competitive Enzyme-linked Immunosorbent Assay (c-ELISA). Overall, seroprevalence of PPR in the area was 36.26% (62/171). All the localities sampled had at least one PPR-positive animal. Age and sex of the animals were not significantly (p>0.05) associated with the infection; however, localities of sampled animals, showed significant (p<0.05) association with PPR virus-infection in goats. It is then concluded that there is high seroprevalence of PPR in traditional raised goats in northern Côte d’Ivoire. Therefore, vaccination campaigns against PPR are advocated to prevent the transmission and spread of PPR in the area.
Sero-epidemiological investigation of foot and mouth disease in cattle at the...ILRI
Poster by Daniel Nthiwa, Silvia Alonso, David Odongo, Eucharia Kenya and Bernard Bett presented at the 15th International Symposium of Veterinary Epidemiology and Ecology, Chiang Mai, Thailand, 12–16 November 2018.
Genomic surveillance of the Rift Valley fever: From sequencing to Lineage ass...ILRI
Poster prepared John Juma, Vagner Fonseca, Samson Limbaso, Peter van Heusden, Kristina Roesel, Bernard Bett, Rosemary Sang, Alan Christoffels, Tulio de Oliveira and Samuel Oyola for the Kenya One Health Online Conference, 6-8 December 2021
Smith TC, Male MJ, Harper AL, Kroeger J, Tinkler G, Moritz-Korolev E, Herwaldt L, Diekema D. High prevalence of MRSA found in Midwestern US Swine and Swine workers. PLoS ONE, 4(1):e4258, 2009.
ABSTRACT- A number of 18 adults male outbred albino rats, weighing between 133-137g were used to investigate the drug susceptibility of Trypanosoma evansi strain isolated from naturally infected dromedary camels in Umbadir area, North Kordofan State, Sudan. The rats were divided into 3 groups (C, D and F) of 6 animals each. Group C and D were infected intraperitoneally with T. evansi (Umbadir stabilate) with 1×104 Trypanosome for the inoculum. Group D rats were given quinapyramine sulphate (20 mg/Kg bwt) after parasitaemia was evident. Group F was left as healthy uninfected control for the stabilate. When parasite counts were one or more parasites per field, counting in haemocytometer were used for exact number of parasite per cubic millimeter using Neubaeur’s counter. Parasites from tail blood were first fixed, stained and diluted in trypanosome diluting reagent. The parasites were diluted to the level that can be easily counted in WBC counting chamber in the haemocytometer. The total number of parasites was expressed as log10 number of parasites per ml of blood. The presence and degree of parasitaemia were determined daily for each rat by examining tail blood. The identity of the local stabilates of Trypanosoma evansi was confirmed through adopting PCR where primers that target the internal transcribed spacer one (ITS1) of the ribosomal DNA were used. There was significant reduction in serum glucose and potassium as well as significant increase in total protein, urea, calcium, albumin and cholesterol in group C. The Umbadir stabilate showed low mortality and high sensitivity to quinapyramine sulphate.
Key-words- Drug susceptibility, T. evansi, Dromedary camels, Sudan
Prevalence of Moraxella ovis Infection in Goats under the Ladang Angkat Progr...iosrjce
IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS) is a double blind peer reviewed International Journal edited by the International Organization of Scientific Research (IOSR). The journal provides a common forum where all aspects of Agricultural and Veterinary Sciences are presented. The journal invites original papers, review articles, technical reports and short communications containing new insight into any aspect Agricultural and Veterinary Sciences that are not published or not being considered for publication elsewhere.
Two main animal pathogenic subspecies of Mycobacterium avium are M. avium avium (Maa) and M. avium paratuberculosis (Map). Their pathogenicity is host-specifi c, Maa causing avian tuberculosis in poultry whereas Map commonly cross-infects to ruminant.Veterinary diagnosis of M. avium infections is microscopic examination of acid-fast bacilli or culture in Löwenstein-Jensen medium,which are time-consuming and low sensitivity. This present study aimed to apply real-time PCR coupled with High-Resolution Melting (HRM) analysis for differential detection of Maa in Thai domestic ducks. Specifi c primer targeting host-expression dependent (hed) region
was designed, PCR product of Maa were amplifi ed from duck’s tissue lesions whereas Map were amplifi ed from cow and deer. HRM real-time PCR was performed and analyzed. Different HRM patterns were showed and melting temperature were analyzed at 83.26 ± 0.12°C for Maa and 84.04 ± 0.09°C for Map. This technique can detect as few as 102 DNA copies and present high specifi city by negative amplifi cation of other pathogenic bacterial species. This technique is sensitive, specifi c, rapid and does not require fl uorescent probes or post-PCR electrophoresis. Our technique is a possible new tool for the detection of Maa and Map infection in tissue specimens.
Two main animal pathogenic subspecies of Mycobacterium avium are M. avium avium (Maa) and M. avium paratuberculosis (Map). Their pathogenicity is host-specific, Maa causing avian tuberculosis in poultry whereas Map commonly cross-infects to ruminant. Veterinary diagnosis of M. avium infections is microscopic examination of acid-fast bacilli or culture in Löwenstein-Jensen medium,which are time-consuming and low sensitivity. This present study aimed to apply real-time PCR coupled with High-Resolution Melting (HRM) analysis for differential detection of Maa in Thai domestic ducks. Specific primer targeting host-expression dependent (hed) region
was designed, PCR product of Maa were amplified from duck’s tissue lesions whereas Map were amplified from cow and deer. HRM real-time PCR was performed and analyzed. Different HRM patterns were showed and melting temperature were analyzed at 83.26 ± 0.12°C for Maa and 84.04 ± 0.09°C for Map. This technique can detect as few as 102 DNA copies and present high specifi city by negative amplification of other pathogenic bacterial species. This technique is sensitive, specific, rapid and does not require fluorescent probes or
post-PCR electrophoresis. Our technique is a possible new tool for the detection of Maa and Map infection in tissue specimens.
Molecular Identification of Bulinus Species in Ogun State, South-West Nigeria...AI Publications
The study considers the distribution of a small sample of 100 Bulinus snails, across 8 localities within Ogun State, Nigerian. Snails were identified using a molecular method of fragment and restriction profiles obtained from ribosomal internal transcribed spacer (its) amplicons. The results showed that the majority of Bulinus samples tested belonged to the species Bulinustruncatus while only one was Bulinusglobosus. The use of Rsa1 restriction endonuclease to cleave the ribosomal its of Bulinus, as a method of species identification, was adopted for the majority of samples, this being a quicker and cheaper method better suited to small laboratory environments. Polymerase chain reaction (PCR) amplification of the schistosome Dra1 repeat within each of the collected Bulinus samples was employed to determine the extent and distribution of infected snails within the sample areas. Successful amplification of the Dra1 repeat demonstrated that 23% of snails were infected with schistosome
Dossier transmission: Transmission of Avian Influenza Virus to DogsHarm Kiezebrink
Avian influenza was found in a dog on a farm in South Gyeongsang Province amid growing concerns that the disease could spread to other animals, officials the Ministry of Agriculture, Food and Rural Affairs said. The dog ― one of three at a duck farm in Goseong-gun, South Gyeongsang Province ― had antigens for the highly pathogenic H5N8 strain of bird flu, the Ministry of Agriculture, Food and Rural Affairs said. The disease affected the farm on Jan. 23.
Since the first case of a dog being infected with the poultry virus in March 2014, there have been 55 dogs found with antibodies to the bird flu virus. The antibody means the immune system of the dogs eliminated the virus. This is the first time bird flu has been found in a dog in Korea through the detection of antigens.
“None of these dogs had shown symptoms. No antigens or antibodies for the virus were found in the two other dogs, which means that dog-to-dog transmission is unlikely to have happened,” quarantine officials said.
The ministry suspected that the dog may have eaten infected animals at the farm. All poultry and dogs at the concerned farm were slaughtered as part of the preventive measures right after the farm was reported to have been infected with the disease, officials said.
Meanwhile, quarantine officials rejected the possibility of viral transmission to humans. According to the ministry’s report, about 450 workers at infected farms across the country had been given an antigen test, with none showing signs of infection. None of Korea’s 20,000 farm workers have reported any symptoms so far, officials added.
“It is thought that infected dogs do not show symptoms of the disease as they are naturally resistant to bird flu,” the ministry said. Meanwhile, the Agriculture Ministry has toughened the quarantine measures in Goseong-gun. The region is a frequented by migratory birds, which are suspected to have spread the viral disease.
African Swine Fever (ASF) virus genomics and diagnosticsILRI
Presented by Richard Bishop and Cynthia Onzere at the Closing workshop of the BecA‐ILRI‐CSIRO‐AusAID project on Understanding ASF epidemiology as a basis for control, Nairobi, Kenya, 2‐3 October 2013
Transmission heterogeneity has consequences on malaria vaccine researches - Conférence du 5e édition du Cours international « Atelier Paludisme » - Vincent ROBERT - Institut de Recherche pour le Developpement, Paris - v.robert@mnhn.fr
"Heart failure is a typical clinical accompanied by symptoms syndrome (e.g. shortness of breath, ankle swelling and fatigue) that lead to structural or functional abnormalities of the heart (e.g. high venous pressure, pulmonary edema and peripheral edema).
In recent years, the significant role of B-type natriuretic peptide has been revealed in the pathogenesis of heart disease and the use of the drug sacubitril/valsartan has started. It has a positive effect on the regulation of the level of B-type natriuretic peptide in the body. It is obviously seen from the the world literature that natriuretic peptides play an important role in the pathophysiology of heart failure. For this reason, many studies suggest that the importance of natriuretic peptides in the diagnosis and treatment of heart failure is recommended.
Due to this, we tried to investigate the effects of a comprehensive medication therapy with a combination of sacubitril/valsartan in the patients with chronic heart failure."
Parallel generators of pseudo random numbers with control of calculation errors
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1. International Journal of Science and Research (IJSR)
ISSN (Online): 2319-7064
Impact Factor (2012): 3.358
Volume 3 Issue 10, October 2014
www.ijsr.net
Licensed Under Creative Commons Attribution CC BY
Prevalence of Canine Parvovirus in Domestic Dogs around Serengeti National Park (Tanzania) Optatus Mwalongo1, Francis Shahada1, Machunde Bigambo2, Paul Gwakisa1, 3, Felix Lankester1, 2, 4 1School of Life Sciences and Bio-Engineering, Nelson Mandela African Institution of Science and Technology (NM-AIST), P.O. Box 447, Arusha, Tanzania 2Serengeti Health Initiative, Arusha, Tanzania 3Genome Science Centre and Dept. of Veterinary Microbiology and Parasitology, Sokoine University of Agriculture, Morogoro, Tanzania 4Paul G. Allen School for Global Health, Pullman, Washington, 99164, USA Corresponding Author: Mobile: +255 759 928 038, E-mail: francis.shahada@nm-aist.ac.tz Abstract: Since 1978 canine parvovirus (CPV) has been an important pathogen of domestic dogs, causing acute haemorrhagic enteritis and myocarditis mostly in puppies. This study was conducted to determine the prevalence of CPV strains in domestic dogs living around the Serengeti National Park (Tanzania). In this study, 77 whole blood samples collected from domestic dogs presenting no clinical signs were tested for CPV antigenic strains (CPV-2a and CPV-2b) using convectional PCR. Eight samples(10.4%) were positive for CPV(CPV-2a and CPV-2b).The study shows that CPV is a common finding in healthy domestic dogs, which suggests they can act as potential reservoirs for transmission to other susceptible domestic dogs and wild life species.
Keywords: canine parvovirus, prevalence, domestic dogs, convectional PCR
1. Introduction
For centuries domestic dogs (Canis familiaris) have been used as human companion animals, hunters and for security purposes, they are one of the most abundant carnivore species [1] and they are found almost everywhere in the world. Since they make up a large contiguous population with many individuals roaming freely [2, 3] this facilitates contact between infected and susceptible individuals, which makes them good reservoirs for pathogens [4] such as canine parvovirus, canine distemper (CDV) and rabies [5]. Canine parvovirus, a small, non-enveloped and single stranded DNA virus belonging to the family parvoviridae [6- 9], emerged in the mid 1970s from felinepanleukopenia virus (FPV)[10-12] when it acquired a new host range in domestic canids. Since then it has caused widespread mortality of domestic dogs [11-14] which, following infection, suffer acute haemorrhagic enteritis and myocarditis [15-17]. The CPV type-2 (CPV-2) which was an original version of domestic dogs evolved from feline panleukopenia (FPV) in the mid 1970s [11, 13, 14] further evolved into three antigenic variants, CPV-2a, CPV-2b and CPV-2c, which subsequently infected, and became endemic in several wild carnivores [8, 18-22]. Although CPV has been reported to affect dogs of all ages (Yang et al., 2010), it primarily affects puppies between six weeks and 4 months of age [22-24]. Following an incubation period from 3 to 5 days [24] the disease manifests itself with fever, vomiting, anoxorea, depression and typical dark or bloody diarrheoa. A blood profile typically shows marked thrombocytopenia and leucopenia [15, 23, 25, 26].
Prevalence of CPV differs from one geographical area to another [27] with the different antigenic variants occurring globally and are found co-circulating [22]. In Africa, CPV strains 2a, 2b and 2c have been reported in several countries including Tunisia, Nigeria, Republic of South Africa, Kenya, Zimbabwe, Zambia, Cape Verde and Namibia [18, 28-31], but there is no available data on the occurrences of CPV in the remaining countries of Africa. However, several of these countries are using the CPV vaccines, suggesting that CPV infections exist in these places too. In most developing countries free-ranging dogs and stray dogs are common, the majority of which are not vaccinated. As the prevalence of CPV tends to be high among unvaccinated dogs [31], it is likely that they serve as reservoirs of the virus [32]. Domestic dogs have been reported to be reservoirs of infectious pathogens and, where they live in close proximity to wild carnivore species, it has been reported that, the spill- over of pathogens from such dogs cause a number of epidemics in wild carnivore species. For instance, they were implicated to be the source of canine distemper epidemic that occurred in Serengeti lions in 1994 and was responsible for nearly 30% deaths the lions [33, 34], rabies epidemic of 1991 in African wild dogs (Lycaon pictus) of the Serengeti ecosystem [35, 36] and they were also associated to be the source of rabies epidemics that affected Ethiopian wolf population [37, 38]. Despite the fact that domestic dogs are implicated as potential reservoirs and source of several infectious diseases including CPV to susceptible wild carnivore species [33, 36], there is no available data on prevalence of CPV in domestic dogs in Tanzania.
Most of the research findings have reported the prevalence of CPV in domestic dogs manifesting clinical signs such as diarrhea, vomiting, depression, dehydration, fever and myocarditis [39], but in this study, the purpose was to
Paper ID: SEP14143 1864
2. International Journal of Science and Research (IJSR)
ISSN (Online): 2319-7064
Impact Factor (2012): 3.358
Volume 3 Issue 10, October 2014
www.ijsr.net
Licensed Under Creative Commons Attribution CC BY
determine the prevalence and types CPV strains in asymptomatic domestic dogs in villages surrounding the Serengeti National Park, Tanzania.
2. Materials and Methods
2.1 Study site The study was carried out in the Mara, Simiyu and Arusha regions of northern Tanzania. 2.2 Samples Collection A total of 77 domestic dog blood samples were used for analysis in this study. The samples were simple randomly selected (in terms of age and location) from an archive that contained frozen samples which had been collected from healthy domestic dogs (both puppies and older dogs had equal chance to be selected for blood sampling) attending rabies vaccination clinics in villages bordering the Serengeti National Park, a zone of 10 Km from the park, between 2007 and 2009 years. The samples used came from 12 villages from Serengeti, Tarime and Loliondo districts found in Mara, Simiyu and Arusha regions, northern Tanzania. 2.3 DNA extraction The viral DNA was extracted from whole blood samples using a ZR Viral DNA Kit (Zymo Research Corp. USA) according to the manufacturer’s instructions. A multivalent vaccine DHLPP (Vanguard, Pfizer-USA) containing attenuated canine parvovirus strain was used as a positive control. 2.4 Primers for convectional PCR The primers used in convectional PCR were designed to amplify VP1/VP2 of the capsid genes. The specific primer pairs; Pbs/Pbas detect CPV type-2b, Pabs/Pabas detect both CPV type-2a and type-2b, and the primer pair H for/Hrev was designed to amplify a larger fragment of DNA for sequencing purposes. The primers Pb and Pab were designed by Pereira et al. (2000) [40] and H was designed by Buonavoglia et al. (2001) [41].The primer pairs Pb and Pab yielded amplicons of the same size, so they were used in separate reactions. All primers used were synthesized by Inqaba biotechnical Industries Ltd (Republic of South Africa) (The primer sequences are shown in Table 1). Table 1: Sequences of the primers used in convectional PCR. s-sense and as-antisense
2.5 PCR assay
The PCR assay was performed as per Pereira et al. (2000) [40] with some modifications. The PCR reaction mixture(25μl) consisted of DreamTaq Green Master Mix(2x) containing: Dream Taq DNA polymerase, 2x Dream Taq Green buffer, dATP, dCTP, dGTP and dTTP,0.4 mM each, and 4 mM MgCl2. The primer pairs used in amplification were Pbs/Pbs, Pabs/Pabs and H for/H rev, each 1μl (0.4μM), 6.5 μl of water free nuclease and 3μl of template DNA. The convectional PCR thermal cycling conditions were set as: activation of DreamTaq DNA polymerase at 94ºC for 3minutes, 30cycles of denaturation at 94ºC for 30 seconds, primer annealing at 55 ºC for 1minute, extension at 72 ºC for 1 minute, final extension at 72 ºC for 5 minutes and 4 ºC final hold. 2.6 Gel-Electrophoresis The Gel-electrophoresis of the PCR products were analyzed using 1.5% agarose gels stained by Gel-Green nucleic acid stain to view the 610bp and 427bp bands so as to verify the presence of the CPV strains . The PCR products were run on gel along with a DNA ladder of 100bp in 1X TBE electrophoresis buffer. Then the results from the gels were visualized by ultraviolet illumination using Gel Doc TM EZ Imager (Bio- Rad Laboratories, Inc) (Figure 1 & 2). 2.7 Statistical Analysis The summarized data were analyzed using Mintab16 (Mintab16, Inc, state college-Pennsylvania) to estimate the CPV DNA prevalence in domestic dogs. The 95% confidence intervals for CPV DNA prevalence were estimated. For this study one sample proportional statistical analysis was performed on Mintab16 platform.
3. Results
The prevalence of CPV DNA in apparently healthy domestic dogs was 10.4 % (8/77) [95% CI=3.58% – 17.22%]. The prevalence of CPV-2a was 6.5% (5/77) and CPV-2b 3.9% (3/77).The CPV positive cases were detected in samples collected from six different villages, namely Merenga (MR), Losoito (LS), Nyamburi(NY), Nyambwaga (NM), Itiryo (IT), and Kitaramanka (KT), in different districts (Serengeti and Tarime). Among the 12 sampled villages, 4 positive cases were from 2 villages in Tarime district (two cases from each, Nyamwaga and Itiryo), also, 4 positive cases were from four villages (one case from each, Bisarara, Merenga, Losoito, and Nyamburi) in Serengeti district, whereas, none of case was detected from among the three sampled villages (Pinyinyi, Engakaseko, and Malambo) in Loliondo district. In summary, Serengeti district-4 cases, Tarime district-4 cases, and none in Loliondo district, therefore, six out of 12 sampled villages had positive cases for CPV-2a and 2b.
The positive samples were from the domestic dogs with the ages of 6, 8, 9 and 10 months, but also 1 and 2 years old [Four cases (50%) were from below one year and the other
Paper ID: SEP14143 1865
3. International Journal of Science and Research (IJSR)
ISSN (Online): 2319-7064
Impact Factor (2012): 3.358
Volume 3 Issue 10, October 2014
www.ijsr.net
Licensed Under Creative Commons Attribution CC BY
four cases (50%) were from one year and above]. In terms of sex, six out of eight of the positive samples were females, furthermore, all the positive CPV cases were from the samples collected in 2008 and 2009 years, and none of the samples from 2007 tested positive. Figure 1: Agarose gel-electrophoresis and GelGreen- florescence of PCR amplified samples using primer Pab. The samples 131,134,135,136,138,139,140 and 106 are positive for CPV-2a/b. Figure 2: Agarose gel-electrophoresis and GelGreen- florescence of PCR amplified samples using primer H. The samples 13, 136, and 107 are positive for CPV.
4. Discussion
This study provides the first data on the prevalence of CPV antigenic strains type-2a and 2b in asymptomatic domestic dogs in Tanzania. Approximately one in ten healthy dogs was positive (8 out 77) which suggests that dogs can be potential carriers of the virus. In this study, the prevalence of antigenic strains CPV-2a was almost twice as high as CPV-2b. This finding is in agreement with studies in countries such as Italy, German, Korea, China, French and Taiwan where CPV-2a has been reported to be more predominant [18, 41-43]. In contrast, CPV-2b has been reported to be the predominant strain in other parts of the world. E.g. Southern Africa, USA, Japan and Turkey [39, 44].The reason why the CPV stains vary in frequencies in different localities is unknown [45]. Since the samples used in this study were collected from only one geographical region (Northern Tanzania including Musoma, Simiyu and Arusha regions), consequently the reported prevalence of CPV-2a is being higher than CPV-b in Tanzania might change if domestic dogs had been sampled from the entire country.
This study reported a prevalence of 10.4% (8/77), but in some other places, there are some reports showing even higher prevalence. For instance, analyses of the faeces of stray dogs in South Korea and Cape Verde (Africa) detected a seroprevalence of CPV-2a of 93.8%, and a prevalence of CPV DNA of 43.3% (23/53) respectively [31, 32]. The reason for variation in proportions of CPV strains in different countries is currently unknown. However, probably this can be associated to poor animal husbandry which normally results in free-ranging/stray dogs and also, ineffective policies for controlling the disease, for example, lack of routine vaccination campaigns. Though, 12 villages were sampled, but the CPV positive cases were detected in samples from only six villages in two districts. In Serengeti district, there were two times the number of villages 4/12 (33%) with positive cases of CPV compared to the number of villages 2/12 (17%) with such cases in Tarime district, and surprisingly, none of the villages in Loliondo district had a positive case. So, Serengeti district had the largest proportion (33%) of the CPV cases detected in domestic dogs followed by Tarime district which had a proportion of 17%. The proportion of the villages without positive CPV-2a and 2b cases from all three districts was 612 (50%). Although, the proportion of the villages with positive cases in Serengeti district was half of the number to that of Serengeti, but there were equal number of the positive cases of the CPV-2a and 2b in such districts. Therefore, from these results we suggest that CPV is localized to some places, since the positive cases were detected in some villages. Normally CPV affects puppies [22, 23], though in some cases it has been reported to affect older dogs [24], but in this study two positive cases were from old domestic dogs with ages above one year. There are two possibilities of explaining this result, in the first case, there is a probability that the positive adult dogs could have been infected when puppies and then remained carriers of the virus, alternatively, the dogs could have been infected at their adult age, supporting the view that the adult dogs are susceptible to canine parvovirus. The detection of canine parvovirus DNA in samples dating back to 2007 shows that CPV has been circulating in domestic dogs for at least seven years. Indeed, 34% (14/55) of sera samples collected from free ranging black-backed jackals (Canis mesomelas) between 1987 and 1988 in Kenya were found to be positive for antibodies against CPV [46] indicating that the virus has been circulating in East Africa since at least 1987. Since only emerged in 1978 in Ontario [47] these findings indicate how fast CPV has spread. The presence of CPV in domestic dogs from the villages around the Serengeti national park, and reports of domestic dogs, which live in villages on the periphery of the Serengeti ecosystem, roaming some distance into park areas in search for food [48] suggests that transmission to wild carnivores is a possibility. Furthermore, transmission might occur when wild carnivores enter human settlement areas to scavenge for food or predate upon domestic dogs [46, 49]. Sick dogs are likely to be easier targets and more frequently eaten which may further potentiate transmission.
Canine parvovirus causes high mortality in puppies dramatically impacting reproductive success [50]. In small and endangered wildlife populations, therefore, the
Paper ID: SEP14143 1866
4. International Journal of Science and Research (IJSR)
ISSN (Online): 2319-7064
Impact Factor (2012): 3.358
Volume 3 Issue 10, October 2014
www.ijsr.net
Licensed Under Creative Commons Attribution CC BY
implications of a CPV outbreak could be catastrophic [51]. Indeed, CPV has been implicated as a barrier to the recovery of wolf populations in North America [52]. The asymptomatic dogs can continue shedding the virus in the environment where it stays viable for longer periods, up to 52 days [16],Therefore, the transmission and spread of the virus can occur in absence of direct contact between the animals, since they can contract it from the contaminated environment [53], this has an implication in making the disease endemic in dogs because other susceptible dogs may contract the virus either from the dog shelter such as kernels or from the contaminated environment.
5. Conclusion
This current study report the detection of CPV-2a and 2b in apparently healthy domestic dogs in regions surrounding the Serengeti ecosystem, Tanzania. The detection of the CPV in asymptomatic dogs shows that the CPV can remain unnoticed among the dog population, hence creates a potential risk in terms of transmission and persistence of the disease, since they can continue shedding the virus in the environment. Furthermore, it was found that CPV-2a is a predominant circulating strain in domestic dogs in Tanzania. Despite of the safe and effective vaccines in place, CPV remains to be an important pathogen which causes mortality and morbidity in domestic dogs in Tanzania. Since the communities living in proximity to the Serengeti ecosystem own domestic dogs which are seldom vaccinated because people are ignorant on the importance of vaccination, poor infrastructures and high costs of the vaccines, these limit the availability of vaccines, consequently, this may result in a large number of unvaccinated dogs of which make the CPV to continue persisting among the dog population, hence serve as reservoirs and source of infections both to other domestic dogs and susceptible wild carnivore species. So vaccination of domestic dog pathogens could probably be an appropriate practice in controlling the disease both in wild and domestic carnivores, so to ensure successes, the practice should be compulsory and enforced by laws.
6. Acknowledgement
The study was financially supported by the Nelson Mandela African Institution of Science and Technology (NM-AIST) and we are so grateful to Carnivore Disease Project (CDP) working under the Serengeti Health Initiative Project (Tanzania) for providing us with samples used in this study. Also, thanks to Nicholaus Peter Myambwa for his wonderful support in technical assistance during my study. References
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Author Profile
Optatus Mwalongo, is a master’s degree student at Nelson Mandela African Institution of Science and Technology in Health and BioMedical Sciences in 2012 and 2014, Arusha, Tanzania. During 2008 and 2011, He pursued his Bachelor of Science in education degree at University of Dar es salaam, Tanzania.
Paper ID: SEP14143 1868