- The document evaluates two autogenous (locally prepared) bacterins against Avibacterium paragallinarum, the cause of infectious coryza disease in chickens.
- 100 layer chickens were divided into 4 groups: two vaccinated groups received bacterins with different adjuvants (aluminum hydroxide or mineral oil), one unvaccinated challenged group, and one unvaccinated unchallenged group.
- After challenge, the vaccinated groups showed fewer clinical signs, deaths, and higher protection rates compared to the unvaccinated challenged group, indicating that both autogenous bacterins were effective at preventing infectious coryza.
Antimicrobial drug resistance pattern of bacteria isolated from cases of abor...Bhoj Raj Singh
Most common causes of abortion and miscarriages include genetic abnormalities in embryo, congenital malformations, immune causes, hormonal causes and infections.
Infections are the least responsible but the most important being extrinsic cause of abortions and thus treatable.
Among the causes of reproductive tract infections commonest are bacteria followed by virus and parasites.
Bacterial infections affect prospective mother right from implantation of the zygote till the postpartum period. Important bacteria are Mycoplasma, Listeria, Salmonella, Brucella and E. coli etc. To treat the bacterial infections antibiotics are the final weapons but proving futile day by day with the emergence of multiple drug resistant (MDR) pathogenic bacteria.
A total of 516 bacterial strains (91 Gram positive and 425 Gram –ve) isolated from cases of abortion (buffaloes 63, cattle 408, goats 14, mares 5, sows 5 and bitches 2) and metritis (bitches 17, buffaloes 2) in animals were tested for sensitivity to:
15 herbal antimicrobials (essential oils of Zanthoxylum rhetsa, Thyme, lemon grass, sandal wood, ajowan, betel leaf, guggul, cinnamon, agar wood, holy basil, patchouli and methanolic extract of Zanthoxylum rhetsa, and three active compounds from herbs viz., carvacrol, cinnamledehyde and citral) and
33 antibiotics (amoxycillin, amoxycillin clavulanic acid, ampicillin, azithromycin, aztreonam cefepime, cefotaxime, cefotaxime clavulanic acid, cefoxitin, ceftazidime, ceftazidime clavulanic acid, ceftriaxone, chloramphenicol, ciprofloxacin, clindamycin, colistin, cotrimoxazole, erythromycin, gentamicin, imipenem, meropenem, moxalactam, nalidixic acid, nitrofurantoin, novobiocin, penicillin, piperacillin, v tazobactam, polymyxin B sulphate , streptomycin, tigecycline and vancomycin) using disc diffusion assay.
To determine extended spectrum β-lactamase and mettalo β-lactamase production specific E-test and polymerase chain reaction assays were performed.
Bacteria belonging to 37 genera were identified from aborted foetal tissues or membranes (497) and bacteria of 6 genera from cases of metritis (19). The 10 most common genera of bacteria associated with abortion were Escherichia (117), Aeromonas (50), Enterobacter (46), Streptococcus (36), Brucella (32), Klebsiella (26), Staphylococcus (22), Alcaligenes (20), Moraxella (19) and Acinetobacter (17).
The bacteria associated with metritis were Staphylococcus (10), Bacillus spp., (2), Enterobacter spp. (1), Escherichia coli (4), Streptococcus milleri (1) and Vibrio alginolyticus (1).
Conclusion: Some of the herbal antimicrobial is as good as antibiotics or even better, the question is how we can use these to alleviate the infections of reproductive tract. In vitro studies are just indicative and real picture may be lucid after in vivo studies for which a concerted one health study is the need of the day.
Emergence of antibiotic resistance in captive wildlifeBhoj Raj Singh
In this presentation antimicrobial drug resistance (AMR) in captive wildlife has been compared with AMR in bacteria isolated from veterinary clinical cases. In captive wildlife bacteria resistant to carbapenems, all generations of cephalosporins, producing ESBL, MBL, and NDM were prevalent. In this study 36.7% bacterial isolates from captive wildlife were ESBL producers and about 45% were MDR type. In recent past not much increase in AMR in bacteria of captive wildlife was observed. Carvacrol was found to be the most effective herbal antimicrobial. About 67.5% bacteria from birds kept in zoo and >71% those from zoo carnivores had MDR. This was much more than in strains of wild herbivore origin (<30%). Herbal drugs resistance was more common in bacteria from herbivore captive wildlife in contrast to AMR for conventional antimicrobials in bacteria from carnivores. Bacteria from carnivores had higher levels of multidrug resistance than those from omnivores or herbivores. No such difference was evident in bacterial isolates from domesticated or pet herbivores and carnivores. Omnivore animals (pigs) and human isolates had almost similar levels of AMR but much higher than herbivores. Position in food chain and Food of the host play an important role in occurrence of AMR bacteria. The study revealed that members of Enterobacteriaceae are the major players in the propagation of AMR. And for maintenance or propagation of AMR Enterobacteriaceae members, wild carnivores are the major abode. The study indicated that AMR was common in bacteria of captive wildlife too as in other biotic and abiotic components of the environment. However, the level of AMR was much more aggravated than in domestic animals. It can be concluded that if we need to monitor the AMR in any locality it will be more informative to look for the AMR strains in wildlife and aquatic environment than in livestock. This might be due to the concentration of the AMR strains in wild carnivores having a specific niche in food chain. More long term studies on large number of isolates from wide variety of captive wildlife living in different geographical and climatological conditions are required for better understanding of AMR trends.
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
All manuscripts are subject to rapid peer review. Those of high quality (not previously published and not under consideration for publication in another journal) will be published without delay.
Antimicrobial drug resistance pattern of bacteria isolated from cases of abor...Bhoj Raj Singh
Most common causes of abortion and miscarriages include genetic abnormalities in embryo, congenital malformations, immune causes, hormonal causes and infections.
Infections are the least responsible but the most important being extrinsic cause of abortions and thus treatable.
Among the causes of reproductive tract infections commonest are bacteria followed by virus and parasites.
Bacterial infections affect prospective mother right from implantation of the zygote till the postpartum period. Important bacteria are Mycoplasma, Listeria, Salmonella, Brucella and E. coli etc. To treat the bacterial infections antibiotics are the final weapons but proving futile day by day with the emergence of multiple drug resistant (MDR) pathogenic bacteria.
A total of 516 bacterial strains (91 Gram positive and 425 Gram –ve) isolated from cases of abortion (buffaloes 63, cattle 408, goats 14, mares 5, sows 5 and bitches 2) and metritis (bitches 17, buffaloes 2) in animals were tested for sensitivity to:
15 herbal antimicrobials (essential oils of Zanthoxylum rhetsa, Thyme, lemon grass, sandal wood, ajowan, betel leaf, guggul, cinnamon, agar wood, holy basil, patchouli and methanolic extract of Zanthoxylum rhetsa, and three active compounds from herbs viz., carvacrol, cinnamledehyde and citral) and
33 antibiotics (amoxycillin, amoxycillin clavulanic acid, ampicillin, azithromycin, aztreonam cefepime, cefotaxime, cefotaxime clavulanic acid, cefoxitin, ceftazidime, ceftazidime clavulanic acid, ceftriaxone, chloramphenicol, ciprofloxacin, clindamycin, colistin, cotrimoxazole, erythromycin, gentamicin, imipenem, meropenem, moxalactam, nalidixic acid, nitrofurantoin, novobiocin, penicillin, piperacillin, v tazobactam, polymyxin B sulphate , streptomycin, tigecycline and vancomycin) using disc diffusion assay.
To determine extended spectrum β-lactamase and mettalo β-lactamase production specific E-test and polymerase chain reaction assays were performed.
Bacteria belonging to 37 genera were identified from aborted foetal tissues or membranes (497) and bacteria of 6 genera from cases of metritis (19). The 10 most common genera of bacteria associated with abortion were Escherichia (117), Aeromonas (50), Enterobacter (46), Streptococcus (36), Brucella (32), Klebsiella (26), Staphylococcus (22), Alcaligenes (20), Moraxella (19) and Acinetobacter (17).
The bacteria associated with metritis were Staphylococcus (10), Bacillus spp., (2), Enterobacter spp. (1), Escherichia coli (4), Streptococcus milleri (1) and Vibrio alginolyticus (1).
Conclusion: Some of the herbal antimicrobial is as good as antibiotics or even better, the question is how we can use these to alleviate the infections of reproductive tract. In vitro studies are just indicative and real picture may be lucid after in vivo studies for which a concerted one health study is the need of the day.
Emergence of antibiotic resistance in captive wildlifeBhoj Raj Singh
In this presentation antimicrobial drug resistance (AMR) in captive wildlife has been compared with AMR in bacteria isolated from veterinary clinical cases. In captive wildlife bacteria resistant to carbapenems, all generations of cephalosporins, producing ESBL, MBL, and NDM were prevalent. In this study 36.7% bacterial isolates from captive wildlife were ESBL producers and about 45% were MDR type. In recent past not much increase in AMR in bacteria of captive wildlife was observed. Carvacrol was found to be the most effective herbal antimicrobial. About 67.5% bacteria from birds kept in zoo and >71% those from zoo carnivores had MDR. This was much more than in strains of wild herbivore origin (<30%). Herbal drugs resistance was more common in bacteria from herbivore captive wildlife in contrast to AMR for conventional antimicrobials in bacteria from carnivores. Bacteria from carnivores had higher levels of multidrug resistance than those from omnivores or herbivores. No such difference was evident in bacterial isolates from domesticated or pet herbivores and carnivores. Omnivore animals (pigs) and human isolates had almost similar levels of AMR but much higher than herbivores. Position in food chain and Food of the host play an important role in occurrence of AMR bacteria. The study revealed that members of Enterobacteriaceae are the major players in the propagation of AMR. And for maintenance or propagation of AMR Enterobacteriaceae members, wild carnivores are the major abode. The study indicated that AMR was common in bacteria of captive wildlife too as in other biotic and abiotic components of the environment. However, the level of AMR was much more aggravated than in domestic animals. It can be concluded that if we need to monitor the AMR in any locality it will be more informative to look for the AMR strains in wildlife and aquatic environment than in livestock. This might be due to the concentration of the AMR strains in wild carnivores having a specific niche in food chain. More long term studies on large number of isolates from wide variety of captive wildlife living in different geographical and climatological conditions are required for better understanding of AMR trends.
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
All manuscripts are subject to rapid peer review. Those of high quality (not previously published and not under consideration for publication in another journal) will be published without delay.
Antibacterial Resistance in the Muscles of Chicken, Pig and Beef IJERA Editor
Though antibiotic drugs are known to improve the health and welfare of food animals , there is parallel risk due
to the development of resistant microorganisms in the body of target animals. Seven meat samples were
procured from wet market in Old Town,Petaling Jaya, Malaysia and assessed for the presence of antibiotic
residues. The samples chosen were chicken parts (skin, muscle and liver) , pig parts (liver, muscle and
intestine) and beef muscle. The results indicated that chicken skin had high level of antibioticresidues which
positively resisted the presence of gram positive, Staphylococcus aureus, S. epidermidisand B. cereus as known
by the zone of inhibition.The beef muscle also held residue which resisted S. aureusChosenbacteriaalong with
the extracts of chicken skin, pig intestine and beef muscle were observed to be resistant totetracycline
hydrochloride, ciprofloxacin hydrochloride monohydrate and their combinations when tested at a concentration
of 1 percent
Strength and weaknesses of fmd control programme going on in india dr. kale b...Bhoj Raj Singh
Foot and Mouth Disease (FMD) is a devastating disease in many of the developing countries including India despite control programs. The FMD in India is associated with loss of about Rs. 20000 crores per annually. Government of India and different provincial governments are spending hundreds of crore rupees per year to control the disease. The FMD control programme (FMD-CP) is running in India since more than 13 years but control of the FMD is still far away dream and the Disease is regularly visiting even the government farms managed by the India's leading Veterinary and Dairy Institutes. The pros and cons of FMD-CP has been discussed in the presentation.
Epidemiologial study of bovine brucellosis in three selected agro-ecologies o...ILRI
Presented by Tujuba Jergefa Oncho at a workshop on an integrated approach to controlling brucellosis in Africa, Addis Ababa, Ethiopia, 29-31 January 2013.
Isolation and Identification of Avibacteriumparagallinarum from Layer Chicken...Dr. Md. Ehsanul Haque
An investigation was conducted for isolation, identification and determination of antibiotic sensitivity of Avibacteriumparagallinarun, the causal agent of infectious coryza, from layer chickens. A total of 21 samples with characteristic symptoms of the disease were collected from a Hatchery of Gazipur. Tissue specimens obtained aseptically from swollen infra orbital sinus and tracheal swab were processed, of which, 3 were found positive while the rest 18 were negative. Isolation of bacteria was performed by first putting the specimen in Nicotinamide adenine dinucleotide (NAD) enriched phosphate buffer broth, anaerobically incubated for 24 hours followed by culturing loopful of broth on Blood agar (BA) and Chocolate agar (CA) plates enriched with NAD and streaked with feeder organism of Staphylococcus. aureus. On 24 hours of anaerobic incubation (candle jar method), dew drop satellite colonies of A. paragallinarum were visible on the culture plates. Cultural characteristics of bacteria as well as their staining, morphological, motility and biochemical properties such as sugar fermentation, MR and V-P tests, Indole production and catalase tests were recorded for identification. Further, antibiogram study revealed that the isolates were sensitive to Ciprofloxacin, Chloramphenicol and Gentamicin but resistant to Ampicillin, Amoxycillin, Oxytetracycline, Erythromycin and Sulphamethoxazole.
Since the European Union’s (EU) ban on the use of Antibiotic growth promoters (AGPs) in animal feed in 2006, the need to find novel strategies to maintain and improve poultry gut health has become ever-important.
Epidemiological research on brucellosis in India: Knowledge generated and gapsILRI
Presentation by Ram Pratim Deka, Johanna Lindahl and Delia Grace at a workshop on One Health approach for brucellosis control in India, New Delhi, India, 26 October 2017.
Production of African Cassava Mosaic Virus (ACMV) Specific Polyclonal Antibod...iosrjce
Serological techniques are commonly used in the detection and characterization of plant viruses.
These methods employ the use of antisera produced by highly purified preparations in intramuscular,
intradermal and intraocular. In this study oral route was explored using crude extracts. Two groups (control
and experimental) of Swiss albino mice consisting of two replicates were immunized via the oral route with
crude extracts from uninfected cassava plants (Manihot esculenta) and cassava plants systematically infected
with African Cassava Mosaic Virus (ACMV). Uninfected and infected leaves were grinded separately in saline
solution (0.15M) at 1:2 (w/v) with laboratory mortar and pestle and then filtered with double layered cheese
cloth of 75µm to obtain extracts. Clarified extracts were orally administered to the mice in daily doses of 200µl
per mice for 21 days and booster doses were also given at day 28 and 35 respectively. Antiserum were obtained
from the mice for 6 consecutive weeks after the commencement of immunization and were analyzed using
antigen coated plate (ACP) and triple antibody sandwich (TAS) indirect enzyme- linked immunosorbent assay
(ELISA). Group A antisera gave negative reactions (OD values < 1.5) while group B antisera reacted positively
(OD values ≥ 1.5) in the two methods used. The polyclonal antisera obtained were very specific to ACMV in
ACP and TAS ELISA. This appears to be the first antisera specific to ACMV obtained by oral immunization of
mice. Oral immunization is considered less stressful for animals, the method is a fast, simple and cheap way for
producing antisera to plant virus compared to the traditional methods of using purified preparations for
immunization. We have used this procedure in the production of antisera yet there is room for improvement in
immunization strategies to enhance antibody production. Immunization dosage can also be tried and
manipulated in bigger animals like rabbits and chicken. This research work leaves room for further exploration
of similar procedure in bigger experimental animals like rabbits and chicken for greater antiserum production.
ABSTRACT- Live microorganisms, have beneficial effects on their host’s health, are called as probiotics. There are various possible sources to isolate
these bacteria. In this studyp harmaceutical probiotic sachet is used as isolation source. The purpose of this study is to search the potentiality of
probiotic bacteria and investigate the probiotic properties of isolates.9 different samples of 3 brands of sachet were used for isolation of bacteria.
Isolates were examined according to their probiotic properties. The probiotic characteristics like pH and Bile tolerance, Antagonistic activity and
Antibiotic susceptibility of isolated bacteria Such as Lactobacillus acidophilus, Lactobacillus rhamnosus and Bifidobacterium bifidum was done. Bile
Tolerance and pH tolerance was determined with the help of the help of coefficient of growth inhibition if their coefficient of growth inhibition is less
than 0.5 the organism was considered as the pH and Bile tolerance. The Strains of Lactobacillus acidophilus and Lactobacillus rhamnosus and Bifidobacterium
bifidum show best result at the pH Acidic to Neutral (5 to 7) and show a bile tolerance from 1-4 % bile. All the isolated bacteria show
the maximum inhibition against Staphyloccocus aureus and minimum against Salmonella typhi by Lactobacillus Strains but Bifidobacterium show
minimum against Escheria coli. Most isolates show resistance toward antibiotics. From this study it can be concluded that pharmaceutical probiotic
products used in the study were showing satisfactory quality and potential probiotic strain.
Key words- Probiotic, Lactobacillus, Bifidobacterium, Sachet
Antibacterial Resistance in the Muscles of Chicken, Pig and Beef IJERA Editor
Though antibiotic drugs are known to improve the health and welfare of food animals , there is parallel risk due
to the development of resistant microorganisms in the body of target animals. Seven meat samples were
procured from wet market in Old Town,Petaling Jaya, Malaysia and assessed for the presence of antibiotic
residues. The samples chosen were chicken parts (skin, muscle and liver) , pig parts (liver, muscle and
intestine) and beef muscle. The results indicated that chicken skin had high level of antibioticresidues which
positively resisted the presence of gram positive, Staphylococcus aureus, S. epidermidisand B. cereus as known
by the zone of inhibition.The beef muscle also held residue which resisted S. aureusChosenbacteriaalong with
the extracts of chicken skin, pig intestine and beef muscle were observed to be resistant totetracycline
hydrochloride, ciprofloxacin hydrochloride monohydrate and their combinations when tested at a concentration
of 1 percent
Strength and weaknesses of fmd control programme going on in india dr. kale b...Bhoj Raj Singh
Foot and Mouth Disease (FMD) is a devastating disease in many of the developing countries including India despite control programs. The FMD in India is associated with loss of about Rs. 20000 crores per annually. Government of India and different provincial governments are spending hundreds of crore rupees per year to control the disease. The FMD control programme (FMD-CP) is running in India since more than 13 years but control of the FMD is still far away dream and the Disease is regularly visiting even the government farms managed by the India's leading Veterinary and Dairy Institutes. The pros and cons of FMD-CP has been discussed in the presentation.
Epidemiologial study of bovine brucellosis in three selected agro-ecologies o...ILRI
Presented by Tujuba Jergefa Oncho at a workshop on an integrated approach to controlling brucellosis in Africa, Addis Ababa, Ethiopia, 29-31 January 2013.
Isolation and Identification of Avibacteriumparagallinarum from Layer Chicken...Dr. Md. Ehsanul Haque
An investigation was conducted for isolation, identification and determination of antibiotic sensitivity of Avibacteriumparagallinarun, the causal agent of infectious coryza, from layer chickens. A total of 21 samples with characteristic symptoms of the disease were collected from a Hatchery of Gazipur. Tissue specimens obtained aseptically from swollen infra orbital sinus and tracheal swab were processed, of which, 3 were found positive while the rest 18 were negative. Isolation of bacteria was performed by first putting the specimen in Nicotinamide adenine dinucleotide (NAD) enriched phosphate buffer broth, anaerobically incubated for 24 hours followed by culturing loopful of broth on Blood agar (BA) and Chocolate agar (CA) plates enriched with NAD and streaked with feeder organism of Staphylococcus. aureus. On 24 hours of anaerobic incubation (candle jar method), dew drop satellite colonies of A. paragallinarum were visible on the culture plates. Cultural characteristics of bacteria as well as their staining, morphological, motility and biochemical properties such as sugar fermentation, MR and V-P tests, Indole production and catalase tests were recorded for identification. Further, antibiogram study revealed that the isolates were sensitive to Ciprofloxacin, Chloramphenicol and Gentamicin but resistant to Ampicillin, Amoxycillin, Oxytetracycline, Erythromycin and Sulphamethoxazole.
Since the European Union’s (EU) ban on the use of Antibiotic growth promoters (AGPs) in animal feed in 2006, the need to find novel strategies to maintain and improve poultry gut health has become ever-important.
Epidemiological research on brucellosis in India: Knowledge generated and gapsILRI
Presentation by Ram Pratim Deka, Johanna Lindahl and Delia Grace at a workshop on One Health approach for brucellosis control in India, New Delhi, India, 26 October 2017.
Production of African Cassava Mosaic Virus (ACMV) Specific Polyclonal Antibod...iosrjce
Serological techniques are commonly used in the detection and characterization of plant viruses.
These methods employ the use of antisera produced by highly purified preparations in intramuscular,
intradermal and intraocular. In this study oral route was explored using crude extracts. Two groups (control
and experimental) of Swiss albino mice consisting of two replicates were immunized via the oral route with
crude extracts from uninfected cassava plants (Manihot esculenta) and cassava plants systematically infected
with African Cassava Mosaic Virus (ACMV). Uninfected and infected leaves were grinded separately in saline
solution (0.15M) at 1:2 (w/v) with laboratory mortar and pestle and then filtered with double layered cheese
cloth of 75µm to obtain extracts. Clarified extracts were orally administered to the mice in daily doses of 200µl
per mice for 21 days and booster doses were also given at day 28 and 35 respectively. Antiserum were obtained
from the mice for 6 consecutive weeks after the commencement of immunization and were analyzed using
antigen coated plate (ACP) and triple antibody sandwich (TAS) indirect enzyme- linked immunosorbent assay
(ELISA). Group A antisera gave negative reactions (OD values < 1.5) while group B antisera reacted positively
(OD values ≥ 1.5) in the two methods used. The polyclonal antisera obtained were very specific to ACMV in
ACP and TAS ELISA. This appears to be the first antisera specific to ACMV obtained by oral immunization of
mice. Oral immunization is considered less stressful for animals, the method is a fast, simple and cheap way for
producing antisera to plant virus compared to the traditional methods of using purified preparations for
immunization. We have used this procedure in the production of antisera yet there is room for improvement in
immunization strategies to enhance antibody production. Immunization dosage can also be tried and
manipulated in bigger animals like rabbits and chicken. This research work leaves room for further exploration
of similar procedure in bigger experimental animals like rabbits and chicken for greater antiserum production.
ABSTRACT- Live microorganisms, have beneficial effects on their host’s health, are called as probiotics. There are various possible sources to isolate
these bacteria. In this studyp harmaceutical probiotic sachet is used as isolation source. The purpose of this study is to search the potentiality of
probiotic bacteria and investigate the probiotic properties of isolates.9 different samples of 3 brands of sachet were used for isolation of bacteria.
Isolates were examined according to their probiotic properties. The probiotic characteristics like pH and Bile tolerance, Antagonistic activity and
Antibiotic susceptibility of isolated bacteria Such as Lactobacillus acidophilus, Lactobacillus rhamnosus and Bifidobacterium bifidum was done. Bile
Tolerance and pH tolerance was determined with the help of the help of coefficient of growth inhibition if their coefficient of growth inhibition is less
than 0.5 the organism was considered as the pH and Bile tolerance. The Strains of Lactobacillus acidophilus and Lactobacillus rhamnosus and Bifidobacterium
bifidum show best result at the pH Acidic to Neutral (5 to 7) and show a bile tolerance from 1-4 % bile. All the isolated bacteria show
the maximum inhibition against Staphyloccocus aureus and minimum against Salmonella typhi by Lactobacillus Strains but Bifidobacterium show
minimum against Escheria coli. Most isolates show resistance toward antibiotics. From this study it can be concluded that pharmaceutical probiotic
products used in the study were showing satisfactory quality and potential probiotic strain.
Key words- Probiotic, Lactobacillus, Bifidobacterium, Sachet
Candida Species As Potential Causes Of Dead–In–Shell Chick Embryos iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
In vitro antimalarial activity of essential oils of Deverra scoparia Coss. & ...INNS PUBNET
The essential oils of the plant Deverra scoparia Coss. & Dur. (Apiaceae) used in traditional medicine in Algeria were subjected to testing the antimalarial activity. Their potential ability to inhibit the in vitro proliferation was evaluated in two strains of Plasmodium falciparum; chloroquine-resistant (K1), and chloroquine-sensitive (3d7) strains. The essential oils displayed good antimalarial activity with IC50 values 1.51 ± 0.71 μg/mL and 0.93 ± 0.89 μg/mL against the 3d7 chloroquine-sensitive and the K1 chloroquine-resistant strains respectively. So, Devera scoparia presented a potential source of antimalarial molecules.
2. Int. J. Poult. Sci., 10 (1): 56-61, 2011
57
MATERIALS AND METHODS
The bacterial strain: The local strain of Avi.
paragallinarum was provided kindly from isolated from
Veterinary Serum and Vaccine Research Institute,
Abbasia, Cairo, Egypt. The strain was isolated from IC
outbreaks in laying flocks then it was identified as Avi.
paragallinarum (Blackall et al., 2005). Also, the
organism was tested to its ability to grow on McConkey
agar, catalase activity, sugar fermentation and its ability
to grow in the presence or absence of Nicotamide
Adenine Dinucleotide (NAD) for the more identification of
the strain (Horner et al., 1992; Mouahid et al., 1992;
Bragg et al., 1993, 1996). This strain was used for
preparation of vaccine, production of antigen for serology
and as challenge bacteria.
The challenge bacterial antigen: For preparation of the
challenge Avi. paragallinarum strain, 7-days old
embryonated chicken eggs was inoculated through the
yolk sac route after Page et al. (1963) and Reid and
Blackall (1983).
Haemagglutinating antigen preparation: The method of
preparation was described after Iritani et al. (1977) and
1980. Strain of Avi. paragallinarum used in agglutination
test was grown in Chicken Meat Infusion (CMI) broth
provided with 0.0025% NAD and incubated at 37 C foro
48 h under microaerophilic conditions (CO2 incubator).
The bacterial antigen was collected from the broth
culture after centrifugation at 6000 r.p.m for 30 h,
washed 3 times in Phosphate Buffer Saline (PBS)
containing 0.01% thiomersal and then the suspension
was titrated and diluted in PBS to provide 4
haemagglutinating units in 0.025% of suspension. The
antigen was stained with crystal violet in concentration
of 0.02%.
Preparation of the autogenous bacterins: A broth
vaccine was prepared from a culture of Avi.
paragallinarum grown in Chicken Meat Infusion (CMI)
broth with 0.0025% NAD and incubated at 37 C for 48 ho
in CO2 incubator (Matsumoto and Yamamoto, 1975;
Davis et al., 1976; Kume et al., 1980). For counting,
aliquot (5 ml of the broth culture) had been removed,
serially diluted in physiological saline and a droplet from
each dilution was placed on CMI supplemented by
0.0025% NAD and incubated under previous conditions.
The number of the Colony Forming Unit (CFU) was
calculated in each dilution. The broth culture was
adjusted to contain at least 10 CFU/ml (Matsumoto and8
Yamamoto, 1975). The culture was inactivated by adding
formalin to a final concentration of 0.25% (Rimler et al.,
1975; Coetzee et al., 1982). Aluminum hydroxide gel
adjuvant was added at concentration of 25% of the final
volume (Matsumoto and Yamamoto, 1971; Boycott et al.,
1977; Blackall and Reid, 1987; Reid and Blackall, 1987;
Blackall et al., 1992) and another bacterin was prepared
by addition of mineral oil (Freund's incomplete adjuvant)
at a concentration of 50% of the final volume (Davis et
al., 1976). Inactivation of bacteria was confirmed by
streaking a 0.1 ml sample on blood and chocolate agar
plates and incubated for 24 h at 37 C in 10% CO2 thato
showed no growth of the bacteria. Both types of
bacterins were stored at 4 C till used.o
Experimental chicks: One hundred, day old LSL layer
pullets were obtained from a commercial hatchery.
Chicks were kept in separate thoroughly cleaned and
disinfected pens and were fed on starter and grower
ration ad libitum. The birds were checked clinically,
bacteriologically and serologically for the absence of Avi.
paragallinarum and other infectious agents just before
grouping or vaccination.
Experimental design: At 6 weeks of age, one hundred
layer chickens were divided into equally distributed 4
groups each containing 25 birds. Group (1) was
vaccinated with Avi. paragallinarum autogenous bacterin
containing aluminum hydroxide, while group (2) received
Avi. paragallinarum autogenous bacterin containing
mineral oil. Both types of bacterins were given in a dose
of 0.5 ml/bird and administered Intramuscularly (IM)
(Blackall and Reid, 1987). Booster dose of both types of
the autogenous bacterins was given in group 1 and 2 at
9 weeks of age. Chickens of group (3) were kept without
vaccination. Birds in groups 1, 2 and 3 were challenged
with 0.2 ml (10 CFU) of live Avi. paragallinarum culture6
by inoculation into the nasal sinus at 12 weeks old
(Yamaguchi et al., 1988). Group (4) was left as blank
control negative (not vaccinated or challenged). Birds
after challenge were kept under complete daily
observation for 7 days.
Criteria for evaluation of the prepared autogenous
bacterins
Signs, mortalities, gross lesions and the protection
rate: All the chickens in the experimental groups were
observed for typical IC signs (nasal discharge, sneezing,
conjunctivitis, swelling of sinuses and facial oedema)
(Yamamoto, 1980) and mortalities daily after challenge
and all the birds were necropsied 7 days after challenge
(end of the study). Chickens were considered protected
if they showed no signs of coryza clinically or at
necropsy.
Bacterial reisolation: Swabs were collected from the
infraorbital sinuses from birds with signs, dead birds as
well as necropsied ones for reisolation of the challenge
organism (Rimler et al., 1975). Each swab sample was
streaked onto CMI agar with 0.0025% NAD and
incubated at 37 C for 48 h in CO2 incubator. Birds wereo
considered protected if they showed no growth of Avi.
paragallinarum on cultures.
3. Int. J. Poult. Sci., 10 (1): 56-61, 2011
58
Serological monitoring: Blood samples were collected challenged groups (1 and 2) at the 4 day after
from ten birds in each group just before vaccination and challenge.
weekly intervals for 3 weeks after vaccination in order to Table 1 showed the efficacy of the two types of
determine the extent of seroconversion of birds to the autogenous Avi. paragallinarum bacterins in layer
bacterins. chickens one week after challenge. It was observed
The agglutination test was conducted as Iritani et al. although there was no significant (p<0.05) differences
(1977) and Thornton and Blackall (1984). Briefly, the between vaccinated groups (1 and 2) but the significant
blood samples were collected from the wing veins of (p<0.05) differences was between the vaccinated and
birds, left to clot, centrifuged and the serum samples the non-vaccinated ones (3 and 4). Chickens vaccinated
were separated. The antibody response was with autogenous aluminum hydroxide adjuvant bacterin
determined by a micro plate agglutination test which showed percentage of clinical signs (8%), dead birds
was performed in 96-well V-shaped micro plates. Serum (4%) and protection rate (92%), while they were 12%, 4%
sample from each chicken was diluted in twofold steps and 88%, respectively in birds vaccinated with
in PBS. Twenty-five micro liters of both Avi. autogenous mineral oil adjuvant bacterin. The
paragallinarum antigen suspension and the serum percentage of clinical signs, dead birds and protection
sample dilutions per well were mixed together and rate were 72%, 16% and 28%, respectively in the
incubated for 18 h at 37 C. The results were recorded as challenged non vaccinated group.o
log2 x of the reciprocal of the highest serum dilution The post mortem lesions of dead as well as sacrificed
which showed agglutination then geometric mean birds 7 days after challenge were catarrhal rhinitis,
antibody titers were calculated. Positive and negative sinusitis and laryngitis with the presence of
sera were included as controls. subcutaneous oedema in the tissues around the
Statistical analysis of the data: The collected data were increase in the vaccinated challenged than the non-
tested using the method of Snedecor and Cochran vaccinated challenged birds.
(1980). The serological results were analyzed by Considering the reisolation rate of Avi. paragallinarum
analysis of variance and by Student’s t-test. Statistical from birds with signs, dead and necropsied ones at the
significance was determined at the 0.05 level of end of the observation period. Table 1 revealed that Avi.
probability. paragallinarum recovered from the infraorbital sinuses
RESULTS
The obtained Avi. paragallinarum strain failed to grow on
McConkey agar and it was catalase negative and
fermented mannitol, fructose, mannose, sucrose,
glucose and sorbitol. Moreover, it failed to produce acid
from arabinose, dulcitol and lactose. The organism
revealed satellites growth on CMI agar and it was NAD-
dependent. From these results, it could be ensure that
the strain was Avi. paragallinarum.
No signs were recorded in the blank non vaccinated 0
challenged group (4). Clinical signs as nasal discharge,
sneezing, conjunctivitis and swelling of the infraorbital
sinuses were observed in non vaccinated and
challenges (group 3) at the 2 day post challenge,nd
whereas signs appeared in the vaccinated and
th
infraorbital sinuses. The severity of lesions was more
of all birds in the non-vaccinated and challenged group
(3), while it was not reisolated from the blank non
vaccinated or challenged group (4). The reisolation rate
of the organism was 12% in chickens received
autogenous aluminum hydroxide adjuvant bacterin and
16% in layers given autogenous mineral oil adjuvant
bacterin.
What was mentioned in Table 2 indicated that there was
gradual increase in the Geometric Mean Titer (GMT) of
antibodies from 3.4 at the 1 week to reach 6.2 log2 atst
the 3 week post vaccination in chickens vaccinated withrd
autogenous bacterin in aluminum hydroxide gel, as well
this titer was elevated from 2.9 to 5.3 log2 at the previous
intervals in layers vaccinated with autogenous bacterin
in mineral oil. Additionally, non-vaccinated chickens did
not develop antibodies.
Table 1: The efficacy of the two types of autogenous Avi. paragallinarum bacterins in layer chickens one week after challenge
Criteria for evaluation
Alum. Mineral ------------------------------------------------------------------------------------------------------------------------
Group No. of hydroxide oil Respiratory Sinus % of % of Protection Reisolation
No. birds adjuvant adjuvant Chall. signs swelling clinical signs dead birds rate (%) rate (%)C C
1 25 + - + 1 1 2/25 (8%) 1/25 (4%) 92% 3/25 (12%)
2 25 - + + 2 1 3/25 (12%) 1/25 (4%) 88% 4/25 (16%)
3 25 - - + 11 7 18/25 (72%) 4/25 (16%) 28% 25/25 (100%)
4 25 - - - 0 0 0/25 (0%) 0/25 (0%) 100% 0/25 (0%)
All the vaccinated groups were differ significantly from the non vaccinated groups (p<0.05). Chall. = Challenge
4. Int. J. Poult. Sci., 10 (1): 56-61, 2011
59
Table 2: The antibody response at the different intervals after vaccination of layer chickens with autogenous Avi. paragallinarum bacterin
Antibody titers at different weeks intervala
Alum. hydroxide Mineral oil --------------------------------------------------------------------------
Group No. adjuvant adjuvant Challenge 1 wk PV 2 wk PV 3 wk PVst nd rd
1 + - + 3.4 5.1 6.2
2 - + + 2.9 4.9 5.3
3 - - + 0 0 0
4 - - - 0 0 0
Wk PV = Week Post Vaccination. Geometric Mean Antibody Titers (GMT) expressed as log2 x (number of birds = 10)a
DISCUSSION
Haemophilus paragallinarum which is now called
Avibacterium paragallinarum (Avi. paragallinarum), the
causative agent of IC in chickens can cause economic
losses to poultry operations throughout the world. The
disease is a highly contagious upper respiratory
disease, characterized by foamy conjunctivitis, sinusitis,
nasal discharges, depression, lethargy and birds may
develop swelling of the wattles and diarrhea. Affected
flocks in lay can suffer drops in egg production from 5 to
10% and in some cases, from 40 to total loss 100% of
eggs (Mouahid et al., 1991). Despite the widespread
use of vaccines against IC, the disease remains a
serious problem in the poultry industry. Several
inactivated vaccines have been used to prevent IC, but
there have been outbreaks reported after using those
vaccines (Davis et al., 1976). It has been well
established that there is no cross protection between
different serogroups (Rimler et al., 1977; Kume et al.,
1980). Thus, the use of autogenous bacterins appears
to be more effective in control of IC if they contain
serovars which are present in a country or locality
(Blackall and Reid, 1987; Mouahid et al., 1991).
Moreover, in order to be successful, these bacterins can
only be used before the birds go into productions, i.e.,
vaccination done at 12-18 weeks of age.
In the present study, double doses of autogenous IC
bacterins (aluminum hydroxide and mineral oil
adjuvanted bacterins) against Avi. paragallinarum was
tested serologically using agglutination test and both
types provoked production of protective immunity in layer
chickens. Similarly, Mouahid et al. (1991) and Glisson
(1998) found that chickens vaccinated with autogenous
bacterin showed a very broad immune response and
protection against Avi. paragallinarum infection.
Previous studies by Otsuki and Iritani (1974) and
Nakamura et al. (1994) reported a close correlation of
antibody titer and protection in chickens vaccinated
against IC. In this work, although the titer of antibodies
was higher in aluminum hydroxide adjuvanted bacterin
than mineral oil one but there was no significant Blackall, P.J., 1988. Further comparison of adjuvants for
difference between them. Davis et al. (1976) reported an inactivated infectious coryza vaccine. Avian Dis.,
accord results. Also, Matsumoto and Yamamoto (1971); 32: 831-835.
Boycott et al. (1977); Kume et al. (1880); Blackall and Blackall, P.J., 1991. An evaluation of the cross-protection
Reid (1987) and Blackall et al. (1992) observed that not afforded by inactivated infectious coryza vaccines.
only IC aluminum hydroxide adjuvanted bacterin was Aust. Vet. Res., 68: 266-267.
protective but also not induce adverse reaction on the Blackall, P.J., 1995. Vaccines against infectious coryza.
vaccination site. World's Poult. Sci. J., 51: 17-26.
The protection rate from IC signs and the clearance of
Avi. paragallinarum from the sinuses were taken as
criteria for evaluation of IC bacterin. By applying these
criteria, we clearly showed that better protection was
obtained when using two shots of the autogenous
bacterins regardless to the type of adjuvant (Yamamoto,
1984; Blackall and Reid, 1987; Blackall, 1988).
Controversial results were obtained by Matsumoto and
Yamamoto (1975); Kume et al. (1980); Coetzee et al.
(1982); Zaki (1985) and Philemon, (2009) who
demonstrated that a single dose of aluminum hydroxide
gel based adjuvant gave protection for layer birds up to
nine months after vaccination. Blackall (1995) concluded
that for longer term protection against IC, aluminum
hydroxide vaccines should be given at two doses, at
least three weeks a part. Autogenous bacterins
containing the prevalent serotypes in an area and
incorporating aluminum hydroxide gel as an adjuvant
appear to be more effective in controlling IC (Reid and
Blackall, 1983; Blackall and Reid, 1987).
Blackall et al. (1992) stated that vaccination programs for
layers and breeders against IC usually consist of a two-
dose regime, the first begins from 6 to 8 weeks of age to
protect pullets against the symptoms, but mainly
function more as a primer for a second vaccination, 6-8
weeks later. For these early vaccinations, poultry
producers prefer the aluminum hydroxide-based
vaccines, due to the minimal local reaction around the
injected site. This reduces the chances of stressing the
pullets at such an important period of feed intake and
proper carcass development. A good vaccination
program with a good primer and booster should provide
long-lasting protection.
In conclusion, both autogenous (aluminum hydroxide
and mineral oil adjuvants) Avi. paragallinarum bacterins
were found to be effective and safe when given as
double I/M shots in layer chickens at 6 and 9 weeks of
age.
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