Intra-species sequence variability in 28s rRNA gene of Oesophagostomum venulosum isolated from goats of West Bengal, India

AbstractObjectiveTo identify genotypes of Oesophagostmum venulosum (O. venulosum) prevailing in West Bengal, India by comparing variation of nucleotide sequences among 28S rRNA.MethodsPCR amplification of partial segment of 28 S rRNA sequence and analysis of sequence amplified product by single strand conformation polymorphism (SSCP).ResultsTwo distinct conformers among male and female parasites were identified by PCR-SSCP analysis. Sequence analysis among conformers revealed the presence of five single nucleotide polymorphisms (SNP) in codon 64, 66, 86, 125 and 146. Secondary RNA prediction structure showed that out of 5 SNPs, 4 occurred at interior loop of RNA which confirmed evolutionary changes among isolates prevailing in this region.ConclusionsSNPs occured in different isolates of O. venulosum might influence critical changes in rRNA folding pattern which influence evolutionary changes among isolates

Izdvajanje, molekularna obilježja i otpornost na antibiotike enterotoksigenih (ETEC) i nekrotoksigenih sojeva E. coli izdvojenih iz zdravih azijskih vodenih bivola.

The present study was undertaken to detect the prevalence, virulence gene profile and antibiotic resistance of Enterotoxigenic Escherichia coli (ETEC) and Necrotoxigenic E. coli (NTEC) in healthy water buffalo in West Bengal. Out of the 363 E. coli isolates from 165 faecal samples, 18 (4.95%) and 7 (1.9%) isolates were found to possess genes for ETEC (LT or STa or STb) and NTEC (cnf1 or cnf2), respectively in PCR. Among the 18 ETEC isolates, 13 (72.22%), 12 (66.67%), and 1 (5.56%) isolates were found to harbour STa, LT, and STb genes respectively. However, among the 7 NTEC isolates, 3 (42.85%) and 4 (57.14%) were detected to possess cnf1 and cnf2 genes, respectively. Further, among the five isolates possessing the EAST 1 gene, four were detected with the STa gene and one (1) with the LT gene. However, the majority of the F41 strains possessed ST genes, and F5 strains harboured both LT and ST. In addition, one out of three cnf1 NTEC isolates was detected to harbour the papC gene and all the four cnf2 NTEC isolates contained the cdt gene. The ETEC isolates belonged to 11 different serogroups (O11, O53, O71, O84, O88, O103, O112, O120, O128, O153, and O158), while, four NTEC isolates belonged to the OUT serogroup and the remaining 3 were disseminated among the O2, O4, and O6 serogroups. The RAPD of the ETEC isolates produced different electrophoretic profifi les. The antibiotic resistance of ETEC and NTEC isolates was observed most frequently to amikacin (56%), kanamycin (44%), gentamicin (40%), neomycin (36%). The study thus revealed water buffalo as a reservoir of multi drug resistant ETEC/NTEC for the first time in India.Istraživanje je provedeno s ciljem da se ustanovi prevalencija, geni za virulenciju i otpornost na antibiotike enterotoksigenih (ETEC) i nekrotoksigenih (NTEC) sojeva bakterije Escherichia coli izdvojene iz zdravih vodenih bivola iz Zapadnog Bengala. Od 363 izolata E. coli iz 165 uzoraka fecesa, 18 (4,95%) je posjedovalo neki od gena za ETEC (LT ili STa ili STb), a sedam (1,9%) izolata imalo je gene za NTEC (cnf1 ili cnf2) dokazane PCR-om. Od 18 izolata ETEC, 13 (72,22%) je imalo gen STa, 12 (66,67%) gen LT i jedan (5,56%) gen STb. Međutim, od sedam NTEC izolata, tri (42,85%) su imala gen cnfl , a četiri (57,14%) gen cnf2. Nadalje, od pet izolata koji su posjedovali gen EAST 1, četiri su imala gen Sta, a jedan gen LT. Većina sojeva F41 posjedovala je gen ST, a sojevi F5 i LT i ST gen. Nadalje, jedan od triju cnf1 NTEC izolata imao je i gen papC, a sva četiri izolata cnf2 NTEC sadržavala su gen cdt. Izolati ETEC pripadali su u 11 različitih seroloških skupina (O11, O53, O71, O84, O88, O103, O112, O120, O128, O153 i O158). Četiri izolata NTEC pripadala su serološkoj skupini OUT, a preostala tri skupinama O2, O4 i O6. RAPD izolata ETEC davao je različite elektroforetske profile. Izolati ETEC i NTEC najčešće su bili otporni na amikacin (56%), kanamicin (44%), gentamicin (40%) i neomicin (36%). Ovim istraživanjem prviput je u Indiji ustanovljeno da su vodeni bivoli rezervoar multiplo rezistentnih sojeva ETEC/NTEC

Prevalencija i otpornost na lijekove šiga-toksigenih (STEC), enteropatogenih (EPEC) i enterotoksigenih (ETEC) izolata bakterije Escherichia coli u slobodno držanih jakova s proljevom i bez proljeva u pokrajini West Kameng, Arunachal Pradesh, Indija.

Food producing animals are the major reservoirs of Shiga-toxin producing (STEC) and enteropathogenic Escherichia coli (EPEC). The yak (Poephagus grunniens) is a unique multipurpose bovid, reared by highlanders in the Himalayan region. A total of 67 STEC, 5 EPEC and 22 ETEC strains were isolated from 256 rectal swabs of free ranging yaks from the West Kameng district of Arunachal Pradesh, India. Among the STEC isolates, shiga toxin producing genes stx2 was predominant, followed by stx1. Of all the stx variants, stx1c, stx2d, stx2c, stx2e and stx2f were detected in 23, 11, 2 and 1 isolates, respectively. Further, genes such as eaeA, ehxA, saa, iha and toxB were detected in 16, 35, 28, 10 and 2 isolates, respectively. One of the EPEC isolates possessed a bfpA gene and was categorized as typical EPEC. Among the ETEC isolates, genes such as LT, STa, STb, F41 and K99 were detected in 14, 5, 13, 2 and 2 isolates, respectively. The majority of the STEC, EPEC and ETEC isolates exhibited multi-drug resistance. The study revealed the presence of multi-drug resistant diarrhoea genic E. coli in free ranging yaks from the West Kameng district of Arunachal Pradesh. Moreover, the presence of STEC/EPEC can be a potential public health risk for tribal highlanders residing in close proximity of the reservoir yaks.Životinje koje služe za proizvodnju hrane mogu biti rezervoar šiga-toksigenih (STEC) i enteropatogenih (EPEC) sojeva bakterije Escherichia coli. Jak (Poephagus grunniens) je jedinstveni multipari bovid kojeg uzgajaju gorštaci na Himalaji. Ukupno je bilo izdvojeno 67 STEC sojeva, 5 EPEC sojeva i 22 ETEC soja iz 256 rektalnih obrisaka slobodno držanih jakova na području West Kameng, Arunachal Pradesh, Indija. Među izolatima STEC prevladavao je gen stx2 za prizvodnju šiga-toksina, a slijedio ga je gen stx1. Od svih varijanata gena stx, stx1c bio je dokazan u 23 izolata, stx2d u 11 izolata, stx2c u dva izolata, a stx2e i stx2f u jednog izolata. Nadalje gen eaeA dokazan je u 16 izolata, gen ehxA u 35, gen saa u 28, gen iha u 10 te gen toxBu dva izolata. Jedan od izolata EPEC posjedovao je gen bfpA i svrstan je u tipične EPEC izolate. Gen LT dokazan je u 14 ETEC izolata, gen Sta u pet, gen STb u 13, gen F41 u dva i K99 također u dva izolata. Većina izolata STEC, EPEC i ETEC pokazivala je otpornost na više lijekova. Istraživanje je pokazalo da postoji višestruka otpornost bakterije E. coli izdvojene iz slobodno držanih jakova s proljevom na području West Kameng, Arunachal Pradesh. Povrh toga, prisutnost STEC/EPEC može predstavljati rizik za javno zdravstvo za gorštačka plemena koja dolaze u bliski dodir s jakovima

Prevalencija i otpornost na lijekove šiga-toksigenih (STEC), enteropatogenih (EPEC) i enterotoksigenih (ETEC) izolata bakterije Escherichia coli u slobodno držanih jakova s proljevom i bez proljeva u pokrajini West Kameng, Arunachal Pradesh, Indija.

Food producing animals are the major reservoirs of Shiga-toxin producing (STEC) and enteropathogenic Escherichia coli (EPEC). The yak (Poephagus grunniens) is a unique multipurpose bovid, reared by highlanders in the Himalayan region. A total of 67 STEC, 5 EPEC and 22 ETEC strains were isolated from 256 rectal swabs of free ranging yaks from the West Kameng district of Arunachal Pradesh, India. Among the STEC isolates, shiga toxin producing genes stx2 was predominant, followed by stx1. Of all the stx variants, stx1c, stx2d, stx2c, stx2e and stx2f were detected in 23, 11, 2 and 1 isolates, respectively. Further, genes such as eaeA, ehxA, saa, iha and toxB were detected in 16, 35, 28, 10 and 2 isolates, respectively. One of the EPEC isolates possessed a bfpA gene and was categorized as typical EPEC. Among the ETEC isolates, genes such as LT, STa, STb, F41 and K99 were detected in 14, 5, 13, 2 and 2 isolates, respectively. The majority of the STEC, EPEC and ETEC isolates exhibited multi-drug resistance. The study revealed the presence of multi-drug resistant diarrhoea genic E. coli in free ranging yaks from the West Kameng district of Arunachal Pradesh. Moreover, the presence of STEC/EPEC can be a potential public health risk for tribal highlanders residing in close proximity of the reservoir yaks.Životinje koje služe za proizvodnju hrane mogu biti rezervoar šiga-toksigenih (STEC) i enteropatogenih (EPEC) sojeva bakterije Escherichia coli. Jak (Poephagus grunniens) je jedinstveni multipari bovid kojeg uzgajaju gorštaci na Himalaji. Ukupno je bilo izdvojeno 67 STEC sojeva, 5 EPEC sojeva i 22 ETEC soja iz 256 rektalnih obrisaka slobodno držanih jakova na području West Kameng, Arunachal Pradesh, Indija. Među izolatima STEC prevladavao je gen stx2 za prizvodnju šiga-toksina, a slijedio ga je gen stx1. Od svih varijanata gena stx, stx1c bio je dokazan u 23 izolata, stx2d u 11 izolata, stx2c u dva izolata, a stx2e i stx2f u jednog izolata. Nadalje gen eaeA dokazan je u 16 izolata, gen ehxA u 35, gen saa u 28, gen iha u 10 te gen toxBu dva izolata. Jedan od izolata EPEC posjedovao je gen bfpA i svrstan je u tipične EPEC izolate. Gen LT dokazan je u 14 ETEC izolata, gen Sta u pet, gen STb u 13, gen F41 u dva i K99 također u dva izolata. Većina izolata STEC, EPEC i ETEC pokazivala je otpornost na više lijekova. Istraživanje je pokazalo da postoji višestruka otpornost bakterije E. coli izdvojene iz slobodno držanih jakova s proljevom na području West Kameng, Arunachal Pradesh. Povrh toga, prisutnost STEC/EPEC može predstavljati rizik za javno zdravstvo za gorštačka plemena koja dolaze u bliski dodir s jakovima

Understanding Antibiotic Usage on Small-Scale Dairy Farms in the Indian States of Assam and Haryana Using a Mixed-Methods Approach-Outcomes and Challenges

The use and misuse of antibiotics in both humans and animals contributes to the global emergence of antimicrobial resistant (AMR) bacteria, a threat to public health and infection control. Currently, India is the world's leading milk producer but antibiotic usage within the dairy sector is poorly regulated. Little data exists reflecting how antibiotics are used on dairy farms, especially on small-scale dairy farms in India. To address this lack of data, a study was carried out on 491 small-scale dairy farms in two Indian states, Assam and Haryana, using a mixed method approach where farmers were interviewed, farms inspected for the presence of antibiotics and milk samples taken to determine antibiotic usage. Usage of antibiotics on farms appeared low only 10% (95% CI 8-13%) of farmers surveyed confirmed using antibiotics in their dairy herds during the last 12 months. Of the farms surveyed, only 8% (6-11%) had milk samples positive for antibiotic residues, namely from the novobiocin, macrolides, and sulphonamide classes of antibiotics. Of the farmers surveyed, only 2% (0.8-3%) had heard of the term "withdrawal period" and 53% (40-65%) failed to describe the term "antibiotic". While this study clearly highlights a lack of understanding of antibiotics among small-scale dairy farmers, a potential factor in the emergence of AMR bacteria, it also shows that antibiotic usage on these farms is low and that the possible role these farmers play in AMR emergence may be overestimated

Antibiotic use, knowledge, and practices of milk vendors in India's informal dairy value chain

BackgroundMilk vendors play an important role in India's dairy value chain; however, their food safety practices are poorly understood. From a milk safety perspective, vendor behavior is significant because it has the potential to affect both consumer and producer behavior. This study describes the types of milk vendors in two Indian states, in an attempt to investigate vendors' hygienic knowledge and practices toward safety and antimicrobial resistance (AMR). MethodsA cross-sectional study was conducted in the states of Assam and Haryana, India. In selected villages, all the milk vendors identified at the time of visit were interviewed. A questionnaire was used to assess the knowledge and practices on antibiotics, milk safety and hygiene. The milk samples were tested for presence of antibiotic resistant bacteria using antibiotic susceptibility testing. ResultsIn total, 244 milk vendors were interviewed during the survey. Out of these, 146 (59.8%) of the vendors traded raw milk, while 40.2% traded pasteurized milk. Vendors were categorized depending on whom they supplied milk to. Five categories were identified: (a) those who sold at grocery shops; (b) those who sold on roadside (roadside vendors); (c) those who sold from door to door; (d) those who sold to sweet makers/tea stalls, and (e) those who sold from own home/other entity. The level of training among vendors on milk hygiene was non-existent and the knowledge related to antibiotics was low. Most of them [210/244 (86.07%)] agreed that boiled milk is always safer than raw milk but almost half [119 (48.77%)] of them admitted that sometimes they drink milk without boiling it. Most vendors believed that they could identify whether milk is safe or not for consumption just by its appearance and smell. Out of 124 milk samples collected from surveyed milk vendors and tested for the presence of antibiotic-resistant bacteria, 80 (64.52%) were tested positive. ConclusionThis study highlights the low levels of knowledge regarding food safety among milk vendors. It shows the predominance of informal milk vendors in the surveyed states and prevalence of AMR bacteria in milk traded by them. Training may be a beneficial strategy for addressing the issue

Antimicrobial resistance pattern, clustering mechanisms and correlation matrix of drug-resistant Escherichia coli in black Bengal goats in West Bengal, India

A cross-sectional study covering four agro-climatic zones of West Bengal, India, was carried out to understand the risk-factors, antimicrobial resistance mechanism and clustering of the resistance characteristics of Escherichia coli isolated from healthy (170) and diarrhoeic (74) goats reared under intensive (52) and semi-intensive (192) farming practices. Of the 488 E. coli isolates, the majority, including the extended spectrum (n: 64, 13.11%) and AmpC β-lactamase (ACBL) (n: 86, 17.62%) producers, were resistant to tetracycline (25.2%), followed by enrofloxacin (24.5%), cefotaxime (21.5%) and amikacin (20.5%). Statistical modelling revealed that the isolates from diarrhoeic animals (p < 0.001) are likely to be more ACBL-positive than those from the healthy counterparts. Similarly, cefotaxime (p < 0.05) and enrofloxacin-resistance (p < 0.01) were significantly higher in diarrhoeic goats and in goats reared intensively. The isolates (n = 35) resistant to multiple drugs revealed the presence of β-lactamase [blaCTXM-1-(21), blaSHV-(7), blaTEM-(3), blaCMY-6-(1), blaCITM-(3)]; quinolone [qnrB-(10), qnrS-(7), aac(6’)-Ib-cr-(3)]; tetracycline [tetA-(19), tetB-(4)] and sulphonamide resistance determinants [sul1-(4)]; multiple plasmids, especially those belonging to the IncF and IncI1 replicon types; and active acrAB efflux pumps. Further, two isolates harbored the carbapenem resistance (blaNDM-5) gene and eight were strong biofilm producers. This first ever study conducted to unravel the status of AMR in goat farming reveals that not only the intensive farming practices but also certain clinical ailments such as diarrhoea can increase the shedding of the drug-resistant isolate. The emergence of multi-drug resistant (MDR) E. coli in goats, particularly those that are carbapenem resistant, is a cause for concern that indicates the spread of such pathogens even in the livestock sub-sector generally considered as naive

Experimental assessment of arsenic toxicity in garole sheep in India

Arsenic, a dangerous bio-accumulative poison, is a grave threat affecting a large number of people as well as animals throughout the World, particularly in Bangladesh and West Bengal, India. It is also a matter of concern as continuously entering into food chain through biotic and abiotic products. The present study was conducted to evaluate the experimental effect of arsenic toxicosis on Garole sheep of West Bengal. One group was subjected to oral arsenic exposure @ 6.6 mg Kg−1 over 133 days when rests considered as negative control. Periodical arsenic estimation in wool, urine and feces along with hemato-biochemical alteration were checked thoroughly. It was evident from the study that long term arsenic exposure exerted a significant (p < 0.01) alteration compared to normal animal which were further supported by clinical abnormalities. Exposed animals showed histological changes throughout major internal organs like coagulative necrosis of liver, tubular nephritis of kidney and acanthosis of skin etc. The bio-accumulative and excretion pattern of arsenic inside body were also well understood by the arsenic estimation study of wool, urine and feces which may be helpful for discussion regarding arsenic entry into food chain via animals

Antibiotic use, knowledge, and practices of milk vendors in India's informal dairy value chain

Background: Milk vendors play an important role in India's dairy value chain; however, their food safety practices are poorly understood. From a milk safety perspective, vendor behavior is significant because it has the potential to affect both consumer and producer behavior. This study describes the types of milk vendors in two Indian states, in an attempt to investigate vendors' hygienic knowledge and practices toward safety and antimicrobial resistance (AMR). Methods: A cross-sectional study was conducted in the states of Assam and Haryana, India. In selected villages, all the milk vendors identified at the time of visit were interviewed. A questionnaire was used to assess the knowledge and practices on antibiotics, milk safety and hygiene. The milk samples were tested for presence of antibiotic resistant bacteria using antibiotic susceptibility testing. Results: In total, 244 milk vendors were interviewed during the survey. Out of these, 146 (59.8%) of the vendors traded raw milk, while 40.2% traded pasteurized milk. Vendors were categorized depending on whom they supplied milk to. Five categories were identified: (a) those who sold at grocery shops; (b) those who sold on roadside (roadside vendors); (c) those who sold from door to door; (d) those who sold to sweet makers/tea stalls, and (e) those who sold from own home/other entity. The level of training among vendors on milk hygiene was non-existent and the knowledge related to antibiotics was low. Most of them [210/244 (86.07%)] agreed that boiled milk is always safer than raw milk but almost half [119 (48.77%)] of them admitted that sometimes they drink milk without boiling it. Most vendors believed that they could identify whether milk is safe or not for consumption just by its appearance and smell. Out of 124 milk samples collected from surveyed milk vendors and tested for the presence of antibiotic-resistant bacteria, 80 (64.52%) were tested positive. Conclusion: This study highlights the low levels of knowledge regarding food safety among milk vendors. It shows the predominance of informal milk vendors in the surveyed states and prevalence of AMR bacteria in milk traded by them. Training may be a beneficial strategy for addressing the issue