The presentation talks about buffaloes' reproduction problems, especially bacterial causes of abortions and uterine infections and their antimicrobial susceptibility.

1. Antimicrobial Susceptibility of Bacterial Causes
of Abortions and Metritis in Buffaloes (Bubalus
bubalis) in Bareilly Region, Uttar Pradesh, India
BR Singh
Pri. Scientist and Head
Division of Epidemiology, ICAR-IVRI,
Izatnagar-243122
Abortions and Metritis in Buffaloes

2. Buffaloes, the poor breeder!!
• In India, ~109.85 million buffaloes (Bubalus bubalis) and ~303
Million cattle.
• Contribute about 55% of total milk production and 30% of the
total meat production in the country.
• Buffalo meat exports account for 89.08% of total animal products
export from India (APFPEDA, 2021).
• Though India holds >58% of the world buffalo population, the
health and production problems of buffaloes are not taken as
seriously as of cattle.
• For economically rearing livestock, productive and reproductive
health is an important determinant.
• Frequent reproductive problems and strong seasonality in
breeding are important reasons for branding buffaloes as poor
breeders (Maherchandani et al., 2018).
• Incidence of reproductive disorders in buffaloes ~37.5% (Kumar et
al., 2015).
Abortions and Metritis in Buffaloes

3. Areas of Major Concern in
Buffalo Production and Reproduction
• Infertility
• Repeat breeding
• Abortion
• Retained placenta, post parturient metritis/ endometritis
• Mastitis
• Calf mortality
• Diseases: HS, BQ, FMD (can modulate sex ratio) and many more
• AI: In 2016: AI service centers- 71,341, 52 million inseminations, success
rate of 35% (Theriogenology. 2016 Jul 1;86(1):194-9. doi: 10.1016/j.theriogenology.2016.04.031.)
• Sexed Semen AI: Pregnancy rates with sexed or non-sexed semen: 38.8%
& 32.3%. doi: 10.1016/j.theriogenology.2011.02.029.
• Quality Semen: Post thaw- there must be >75% live and normal sperms
and forward motility should be >70% but in a sample survey of sexed
semen at College of Veterinary Science, Meerut, only 28% live sperms and
20% motility was detected, still then the quality certified (an RTI).
Abortions and Metritis in Buffaloes

4. Origin of Reproductive problems
• Genetic
• Anatomical
• Physiological
• Nutritional
• Managerial: Including AI and natural breeding
• Infectious: Bacterial, Viral, Fungal, Protozoal,
Rickettsial, Mycoplasmal, mixed (multiple).
Abortions and Metritis in Buffaloes

5. Common Bacterial Causes of Abortions in
Buffaloes, reported earlier
• Abortions in buffaloes are common and the
incidence may vary from 1.51% to 7.1% in
different breeds (Galiero, 2007; Satish et al.,
2019).
• Brucella spp., Arcanobacterium pyogenes,
Chlamydophila spp., Coxiella burnetii, Bacillus
licheniformis, Campylobacter fetus venerealis,
E.coli, Leptospira spp., Listeria monocytogenes
Abortions and Metritis in Buffaloes

6. Common causes of uterine infection in
buffaloes, reported earlier
• Rao (1982) ~30% incidence of endometritis, Kumari et
al. (2016) reported it ~9.66 % in India.
• Moghaddam and Mamoei (2004) incidence of 29.4% of
infertility and endometritis in Iran.
• Ahmad et al. (2008), the incidence of endometritis in
Egyptian buffaloes ~14.10%.
• Durani et al. (2009), 12.8% of buffaloes in Pakistan had
uterine infections.
• E. coli, Arcanobacterium pyogenes, Bacteroides fragilis,
Fusobacterium necrophorum, Streptococcus spp.,
Staphylococcus spp., and Pseudomonas spp.
Abortions and Metritis in Buffaloes

7. This study
• 23 Aborted foeti’s stomach contents.
• 20 Uterine aspirates from uterine infection cases.
• Anaerobic, Aerobic and Microaerobic Culture (Carter et al., 1975).
• Antibiotic susceptibility assay (CLSI, 2014, 2015).
• Herbal antimicrobial drug susceptibility assay (Singh, 2013).
• Detection of antibodies in serum of aborted buffaloes by Standard Tube
agglutination assay (Carter et al., 1975).
Abortions and Metritis in Buffaloes

8. Gram-ve Bacteria isolated from aborted foeti
• Achromobacter xyloxidans 1,
• Acinetobacter calcoaceticus 1, A. schindleri 1,
• Aeromonas bestiarum 1, A. media 2,
• Alcaligenes denitrificans 1, A. faecalis 6,
• Brucella abortus 4,
• Edwardsiella tarda 1,
• Enterobacter agglomerans 4, E. amnigenus 1,
• Pectobacterium cyperipedii 1,
• Escherichia coli 8,
• Hafnia alvei 1,
• Klebsiella pneumoniae ssp. pneumoniae 4,
• Moelerella wisconsensis 1,
• Pseudomonas testosteronii 1,
• Salmonella enterica ser Dublin 1,
• Xenorhabdus bovienii 2, X. poinarii 1
Abortions and Metritis in Buffaloes

9. Gram+ve Bacteria isolated from aborted
foeti
• Aerococcus spp. 1,
• Bacillus subtilis 1,
• Enterococcus avium 1, E. durans 1,
• Kocuria varians 3,
• Staphylococcus aureus 1, S. warneri 1,
• Streptococcus agalactiae 1, S. bovis 1, S. uberis
Abortions and Metritis in Buffaloes

10. Antibodies to isolated bacteria in aborted
buffalo serum
Bacteria Case
Number
Agglutination
titres
Salmonella enterica ser Dublin 2 40
Brucella abortus 3 >320
Brucella abortus 4 >320
Alcaligenes faecalis 5 >320
Escherichia coli 6 >320
Escherichia coli 7 >320
Escherichia coli 8 >320
Klebsiella pneumoniae 9 80
Enterobacter agglomerans 10 40
Enterobacter agglomerans 11 >320
Klebsiella pneumoniae 12 40
Enterobacter amnigenus 14 80
Xenorhabdus poinarii 15 >320
Aeromonas media 17 160
Brucella abortus 18 >320
Brucella abortus 19 >320
Abortions and Metritis in Buffaloes

11. Gram-ve Bacteria isolated from uterine
infection cases
• Acinetobacter lowffii 6,
• Aeromonas bestiarum 1, A. salmonicida 1,
• Alcaligenes denitrificans 2,
• Enterobacter agglomerans 2,
• Escherichia coli 9,
• Hafnia alvei 1,
• Proteus vulgaris 4,
• Vibrio alginolyticus 1
Abortions and Metritis in Buffaloes

12. Gram+ve Bacteria isolated from uterine
infection cases
• Bacillus pantothenticus 2,
• Enterococcus faecalis 1, E. faecium 2,
• Kocuria varians 3,
• Staphylococcus intermedius 1,
• Streptococcus morbilorum 1, S. pneumoniae 1
Abortions and Metritis in Buffaloes

13. Bacteria with superbug potential from
abortion cases
MBL Bacteria from abortion cases
• Enterobacter amnigenus,
• Aeromonas bestiarum,
• Hafnia alvei,
• Acinetobacter calcoaceticus,
• Moellerella wisconsensis,
Carbapenem resistant non-MBL Bacteria
• Aeromonas media
• Staphylococcus warneri
Abortions and Metritis in Buffaloes

14. Bacteria with superbug potential from
uterine infections
MBL Bacteria
• Aeromonas salmonicida
• Alcaligenes denitrificans
• Acinetobacter lowffii (from 5 cases)
• Escherichia coli (from 2 cases)
Carbapenem resistant non-MBL Bacteria
• Kocuria varians
• Enterococcus faecium
• Escherichia coli
Abortions and Metritis in Buffaloes

15. Most effective to the least effective (% susceptible)
herbal antimicrobials on bacteria associated with
abortion and metritis in buffaloes
On abortion causing bacteria On metritis causing bacteria
Ajowan oil 100.00 Carvacrol 98.53
Thyme oil 98.18 Cinnamledehyde 97.37
Cinnamledehyde 96.36 Ajowan oil 92.11
Cinnamon oil 96.36 Thyme oil 89.47
Carvacrol 95.29 Cinnamon oil 84.21
Holy basil oil 90.59 Holy basil oil 79.41
Citral 61.43 Citral 69.81
Sandalwood oil 38.57 Lemongrass oil 60.53
Lemongrass oil 36.36 Sandalwood oil 45.28
Agarwwod Oil 25.71 Betel leaf oil 42.11
Betel leaf oil 20.00 Agarwwod Oil 41.51
Patchouli essential oil 9.09 Patchouli essential oil 23.68
Guggul oil 5.45 Guggul oil 18.42
Abortions and Metritis in Buffaloes

16. Ten most effective (% susceptibility)
antibiotics on bacteria isolated from
abortion and metritis cases in buffaloes
On bacteria from aborted foeti On bacteria from uterine infections
Tigecycline 98.18 Tigecycline 94.74
Ceftriaxone 89.09 Chloramphenicol 92.11
Imipenem 89.09 Cefepime 78.95
Meropenem 89.09 Meropenem 73.68
Colistin 88.37 Gentamicin 71.05
Chloramphenicol 83.64 Imipenem 71.05
Gentamicin 81.82 Moxalactam 63.16
Azithromycin 80.00 Tetracycline 63.16
Cefotaxime 80.00 Cefotaxime 60.53
Cefepime 78.18
Ampicillin+sulbactam 57.89
Abortions and Metritis in Buffaloes

17. Ten least effective (% susceptibility) antibiotics on
bacteria isolated from abortion and metritis cases
in buffaloes
On bacteria from aborted foeti On bacteria from uterine infections
Erythromycin 18.18 Novobiocin 18.18
Penicillin 25.45 Amoxicillin 18.42
Ampicillin 29.09 Erythromycin 23.68
Amoxicillin 32.73 Penicillin 28.95
Novobiocin 33.33 Cotrimoxazole 31.58
Ceftazidime 45.45 Vancomycin 36.36
Amoxicillin+clavulanic
acid
50.91 Ampicillin 36.84
Aztreonam 53.49 Ceftazidime 36.84
Ampicillin+sulbactam 60.00 Amoxicillin+clavulanic
acid
47.37
Vancomycin 66.67 Piperacillin 47.37
Abortions and Metritis in Buffaloes

18. Conclusions
• No fixed set of bacteria causing metritis and abortions in buffaloes.
• Antimicrobial susceptibility of bacteria not only depends on the type of
bacteria but on the problem they are associated with.
• Bacteria isolated from cases of uterine infections are more often (p, 0.01)
resistant to multiple antibiotics than those associated with abortion.
• Although a few of the antibiotics were comparatively more effective are
not recommended for animal use.
• Antimicrobial susceptibility testing of the isolates should always be
recommended to determine the therapeutic usefulness of antimicrobials
to be used.
• Uterine tract infections are usually chronic and the treatment can be
deferred for a few days to get the antimicrobial susceptibility report
rather than wasting valuable antimicrobials and precious breeding time
of buffaloes.
• Although herbal antimicrobials may be tried as antibiotic alternatives,
studies on suitable formulations are needful for conformity in the
therapeutic outcome.
Abortions and Metritis in Buffaloes

19. Abortions and Metritis in Buffaloes