Detection of bovine respiratory syncytial virus using a heterologous antigen-capture enzyme immunoassay

Based on the marked antigenic similarities that exist between antigens of the human and bovine strains of respiratory syncytial virus (RSV), an enzyme immunoassay (EIA) designed to detect human RSV was used to detect bovine RSV. The commercial test kit (RSV EIA) consists of a solid phase (beads) coated with a capture antiserum prepared against the Long strain of human RSV. The RSV EIA test was compared with the method of inoculation of cell cultures and fluorescent antibody (FA) staining of lung tissue for the detection of bovine RSV. Using a cell culture-propagated stock of strain 375 of bovine RSV, the threshold of sensitivity of the EIA test for the cattle strain of RSV was determined to be less than or equal to 10(2.3) CCID50/ml. In addition, RSV EIA detected the bovine RSV in nasal samples obtained from 3 experimentally inoculated cattle. The RSV EIA exhibited a sensitivity of greater than or equal to 80% during the period that shedding of infectious virus took place. All of the bovine RSV FA-positive lung samples (n = 37) were positive by the RSV EIA. Twenty-six of the remaining 214 bovine RSV FA-negative lung samples were positive by the RSV EIA. The RSV EIA was also used to test 137 nasal swabs obtained from cases of bovine respiratory disease. Of these, 38 tested positive by RSV EIA. All samples that tested positive by EIA were confirmed by blocking assays using hyperimmune serum anti-bovine RSV and a pool of monoclonal antibodies specific for that virus.     

Validation of synthetic peptide enzyme immunoassays in differentiating two subgroups of ruminant respiratory syncytial virus.

Subgroup-specific peptide-based enzyme immunoassays from each respective G-glycoprotein of the ovine and the bovine respiratory syncytial virus (RSV) were developed to detect RSV-specific IgG responses in cattle. Antigenic peptides from the respective G-glycoprotein were identified from the extracellular central hydrophobic region (amino acids 158-189) located between 2 mucin-rich regions. These antigenic peptides identified by epitope mapping from each G-glycoprotein were synthesized and used to develop the subgroup-specific enzyme immunoassays. The negative cutoff for each enzyme immunoassay was established as the mean optical density of indirect immunofluorescent antibody-negative bovine sera plus 3 SDs. The sensitivity (82.9%) and specificity (100%) of the bovine enzyme immunoassay and the specificity (95.8%) of the ovine enzyme immunoassay were determined by comparison with indirect immunofluorescence (used as the "gold standard"). The negative and positive predictive values were calculated for each assay. The presence of serum antibody to ovine RSV in cattle implies that this virus infects cattle and may contribute to the pathogenesis of bovine respiratory disease.     

A preliminary serological survey of viral antibodies in Peruvian sheep

This study reports the sero-prevalence of viral infections in sheep in Peru. Serum samples were collected from 34 mature healthy rams located in 3 different geographic regions of the country (north, central and south). The sera were tested for antibodies to the following viruses: respiratory syncytial virus (RSV); parainfluenza 3 (PI-3) virus; bovine viral diarrhea/border disease (BVD/BD) virus; bovine herpesvirus 1 (BHV-1); bluetongue (BT) virus; ovine progressive pneumoniae (OPP) virus; bovine leukosis virus (BLV). The serological studies showed that 47% were positive for RSV; 82% for PI-3; 3% for BVD/BD virus; 49% for BT virus; 13% for OPP virus. Antibodies were not detected to bovine herpesvirus 1 or to bovine leukosis virus.     

Genetic characterisation of antibiotic resistance and virulence factors in vanA-containing enterococci from cattle, sheep and pigs subsequent to the discontinuation of the use of avoparcin

The prevalence of vancomycin resistant-enterococci (VRE) in faecal samples from cattle, sheep and pigs slaughtered for human consumption was evaluated. Enterococci containing the vanA gene were detected in 25.3% and 2.7% of the porcine and ovine samples, respectively, and were identified as Enterococcus faecium. No vanA-containing enterococcal strains were detected in bovine samples. Enterococcal strains with intrinsic vancomycin resistance were detected in seven (9.9%) faecal samples from pigs and in two samples from both cattle and sheep (3.7% and 2.7%, respectively). All vanA-positive isolates from pigs were resistant to tetracycline and erythromycin, and the mobile element Tn916/Tn1545-like transposon was detected in 90.5% of the tetracycline-resistant isolates that contained the tet(M) gene. Although gelatinase and haemolytic activity were not detected, the hyl and cylB virulence genes were found within the VRE strains isolated.     

Differentiation between bovine and ovine strains of Histophilus somni based on the sequences of 16S rDNA and rpoB gene

Nucleotide sequences of 16S rDNA and rpoB gene of 25 bovine and 6 ovine Histophilus somni strains were determined to detect subtle differences between the host animal species. The 1465 nucleotide residues of the 16S rDNA exhibited levels of sequence similarities of 99.4% or more. The high sequence similarity of the 16S rDNA of recently described species H. somni was confirmed in the 31 strains from cattle and sheep. These results suggested that the intra-specific diversity of 16S rDNA was limited in bovine and ovine strains of H. somni. The specific association of strains was also observed in the 311 bp region of rpoB gene which sequence similarities were 98.6% or more. However, the phylogenetic tree analysis of the rpoB gene showed that the ovine strains appeared to form a subgroup recovered in 70% of the bootstrap trees. In the 311 bp region of the ovine strains, a HincII restriction endonuclease site was detected. The PCR-amplified rpoB DNA of 46 bovine and 20 ovine H. somni strains were examined for the digestion with HincII. As the results, 17 strains of ovine strains were cleaved by the enzyme but none of the bovine strains appeared to possess the restriction site. The restriction enzyme analysis of rpoB gene may be useful to differentiate ovine strains from bovine strains of H. somni.     

Peste des petits ruminants infection in domestic ruminants in Sudan

The existence of peste des petits ruminants (PPR) in domestic ruminants and camels in Sudan during 2008–2012 was investigated. Lung tissues and serum samples were randomly collected from sheep, goats, cattle, and camels at different areas of Sudan. A total of 12,384 serum samples were collected from clinically healthy 7413 sheep, 1988 camels, 1501 cattle, 1459 goats, and 23 gazelles at different areas in the Sudan. They were examined for PPR antibodies using competitive ELISA (cELISA). The overall detected seroprevalence of PPR in tested sera was 49.4%; seroprevalence values within species were 67.1, 48.2, 25.8, 2.1, and 21.7% in sheep, goat, cattle, camels, and gazelles, respectively. The highest seroprevalence (68.1%) was observed in sera collected from Darfur states, then the central states (54.3%). A total of 1276 lung tissue samples (623 sheep, 324 cattle, 220 camels, and 109 goats) were collected. The majority of lung samples were collected from clinically healthy animals that showed lesions on PM in slaughterhouses (95%) and during PPR outbreaks; samples were tested for PPR antigen using immunocapture ELISA (IcELISA). PPR antigen was detected in 233 out of the 1276 tested samples (18.3%). Positive results were observed in samples collected from clinically healthy and diseased animals. The observed prevalence values in each species were 33.6, 21.1, 15.4, and 12.3% in camel, goat, sheep, and cattle, respectively. PPR antigen was detected in samples from different areas; however, the highest prevalence (63.9%) was found in samples collected from the eastern states, then Khartoum state (28%). Trials for virus isolation were done in different cell cultures. Out of 30 IcELISA-positive samples inoculated in primary bovine and ovine kidney cells, Vero cells, the PPR virus was successfully isolated from 15 (eight sheep, five camels, and two goats) samples in the three cell culture types. Using RT-PCR, PPRV nucleic acid was detected in all 25 IcELISA-positive tested samples.     

Mass screening of cattle sera against 14 infectious disease agents, using an ELISA system for monitoring health in livestock.

Mass screening ELISA methods were developed for testing cattle serum for antibodies against 14 common livestock diseases simultaneously. The absorbance values were transformed to a %ELISA (spectrophotometric antibody end point) by a computer interfaced with a microplate reader. A histogram indicating a cutoff point and a report for the veterinarian also was generated. The computer program produced a print-out of the antibody profile for each animal tested, the antibody concentration against each disease, and a histogram (antibody profile) showing the prevalence of each disease in the herd. Serum samples were obtained from 1,953 cattle, including 880 dairy cattle from 10 herds and 1,073 beef cattle from 20 herds. These samples were obtained from June 1988 through June 1989. The highest antibody prevalence was against bluetongue virus. Of the 1,953 cattle tested, 1,223 (63%) were seropositive for bluetongue virus, including 502 (57%) of the dairy cattle and 721 (67%) beef cattle. Other antibody prevalences, in descending order, were: rotavirus (44%), Pasteurella spp (25%), Leptospira spp and Haemophilus spp (22%), Mycoplasma spp (18%), parainfluenza virus (17%), Campylobacter spp (16%), Anaplasma marginale (15%), bovine leukosis virus (13%), Brucella spp (8%), Mycobacterium paratuberculosis (8%), bovine viral diarrhea virus (3%), and infectious bovine rhinotracheitis virus (3%). Major differences in antibody prevalence between dairy and beef cattle were that only 4% of the dairy cattle were seropositive for A marginale, compared with 25% of the beef cattle, and conversely, 29% of the dairy cattle were seropositive for bovine leukosis virus, compared with 1% of the beef cattle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Milk whey induction of agglutination in ovine and bovine mastitis Staphylococcus aureus

A total of 59 mastitis staphylococcic strains were tested for growth agglutination upon supplementation of growth media with ovine and bovine milk whey and mammary secretions from dry cows. Differences were observed when comparing bacterial species or origins (ovine vs. bovine) of bacteria and whey. All of the ovine and bovine S. aureus strains tested, but only 4 among 22 other ovine mastitis staphylococcic strains, showed growth agglutination in Todd Hewitt broth (THB) supplemented with greater than or equal to 30% (v/v) ovine milk whey. None of the strains agglutinated during growth in regular THB medium. Ovine whey had an agglutination induction capacity higher than bovine whey (P less than 0.005), concerning the number of responsive ovine and bovine S. aureus strains. There were no differences between whey samples from different ewes with regard to their capacity to induce agglutination. Ovine S. aureus strains were more responsive than bovine strains of this bacterial species, concerning the number of responsive strains (P less than 0.001) to bovine whey (greater than or equal to 30% in THB), the proportion of responsive strains at low (10%) ovine whey concentration (P less than 0.001), and the strength of reaction (precipitation timing and clump size). Secretions from dry cows systematically induced agglutination in all of the bovine and ovine S. aureus strains tested.     

Genetic analysis of the G and P genes in ungulate respiratory syncytial viruses by RNase A mismatch cleavage method

The G and P genes of bovine, ovine and caprine respiratory syncytial (RS) viruses were analyzed by RNase A one-dimensional fingerprinting, using A51908 as the reference strain. Antisense G or P RNA probes of bovine RS virus strain A 51908 were hybridized to total RNA extracted from bovine turbinate cells infected with bovine, ovine or caprine RS virus strains. The RNA:RNA heteroduplexes were digested with RNase A and the resistant products were analyzed by gel electrophoresis. Comparative analysis of the cleavage patterns revealed heterogeneity among bovine, ovine and caprine RS virus isolates. Ovine RS virus strains generated RNA cleavage patterns more distantly related to the bovine or caprine RS virus strains, particularly in the G gene. Statistical analysis of the results obtained indicated that genetic differences between bovine and ovine viruses were larger, compared with the ones among bovine strains themselves. The same analysis also revealed a close genetic relation among bovine and caprine strains. These results are discussed in terms of ungulate RS virus genetic variation and vaccine development.     

Prevalence of antibodies to seven viruses in a flock of ewes in Minnesota

Blood samples were collected from a flock of healthy ewes at a University of Minnesota research station. Sera from these blood samples were tested for antibodies against 7 viruses, using 3 tests (eg, virus-neutralization test for bovine viral diarrhea virus, infectious bovine rhinotracheitis virus, bovine adenovirus type 3, and bovine respiratory syncytial virus; hemagglutination inhibition test for parainfluenza virus type 3; and agar-gel immunodiffusion test for lentivirus of ovine progressive interstitial pneumonia and bluetongue virus). The number of seropositive ewes for each antibody type were 1 of 377 (0.3%) for bovine viral diarrhea virus, 2 of 377 (0.5%) for infectious bovine rhinotracheitis virus, 29 of 378 (7.6%) for bovine adenovirus type 3, 200 of 378 (52.5%) for bovine respiratory syncytial virus, 273 of 373 (71.7%) for parainfluenza virus type 3, and 210 of 379 (55%) for ovine progressive pneumonia virus. All ewes were seronegative for bluetongue virus antibodies.     

A mixed methods study of ruminant brucellosis in central-eastern Tunisia

In this study, we conducted an investigation to determine the true prevalence of bovine and ovine brucellosis in central-eastern Tunisia. A total of 1134 veterinary samples taken from 130 ruminant herds were screened for brucellosis using IS711-based real-time PCR assay. Sera collected from the ruminants were tested using the Rose Bengal test and indirect enzyme-linked immunosorbent assay. Based on serological and molecular results, the true adjusted animal population level prevalence was 23.5 % in cattle, against 13.5 % in sheep. In addition, the true adjusted herd level prevalence of brucellosis was 55.6 % in cattle and 21.8 % in sheep. A statistically significant association was found between vaginal and milk shedding for ruminants. In addition, our results showed that Brucella abortus could be responsible for bovine and ovine brucellosis. Multivariable logistic regression analysis at the animal population level indicated that age and origin variables were important risk factors for cattle. However, age and abortion variables were found to be associated with ovine brucellosis. At the herd level, risk factors for Brucella positivity were as follows: abortion and herd composition for cattle against herd composition, mortality rates, and hygiene for sheep. Animal hygiene, food quality, and sanitary practices on the farm should be applied as strategies to control brucellosis in herds.     

Genetic diversity and frequency of bovine viral diarrhea virus (BVDV) detected in cattle in Turkey

The aim of this study was to investigate the frequency and diversity of bovine viral diarrhea viruses (BVDV) infecting cattle in Turkey. A total of 1124 bovine blood samples from 19 farms in 4 different Turkish regions were tested by antigen capture ELISA (ACE). BVDV antigen was found in 26 samples from 13 farms. Only 20 of the 26 initial test positive cattle were available for retesting. Of these, 6 of 20 tested positive for BVDV, by ACE and real-time RT-PCR, one month after initial testing. Phylogenetic analysis, based on comparison of the E2 or the 5'UTR coding regions, from 19 of the 26 initial positive samples, indicated that 17 belonged to the BVDV-1 genotype and 2 to the BVDV-2 genotype. Comparison of 5'UTR sequences segregated 8 BVDV-1 strains (strains 5, 6, 10, 11, 12, 13, 17, and 19) to the BVDV1f, 1 strain (strain 8) to the BVDV1i and 1 strain (strain 14) to the BVDV1d subgenotypes. One strain (strain 4) did not group with other subgenotypes but was closer to the BVDV1f. The remaining 6 BVDV-1 strains (strains 1, 2, 3, 7, 9, and 18) segregated to a novel subgenotype. The E2 sequence comparison results were similar, with the exception that strain 5 grouped with the novel subgenotype rather than BVDV1f subgenotype. It appears that among the diverse BVDV strains in circulation there may be a subgenotype that is unique to Turkey. This should be considered in the design of diagnostics and vaccines to be used in Turkey.     

1株牛病毒性腹泻病毒分离毒株的基因组特征

旨在从宁夏某奶牛群持续感染牛分离牛源牛病毒性腹泻病毒（BVDV）,并解析其基因组特征,为研究我国不同地区BVDV分离株遗传演化规律提供理论依据。利用BVDV抗原检测试剂盒检测宁夏回族自治区银川市某示范区的240头高产奶牛间隔两周的双份抗凝血,筛选持续感染牛,分离血液淋巴细胞制备裂解液接种牛肾细胞（MDBK）,分离鉴定获得BVDV株,克隆测序获得全基因组序列,比较分析其遗传演化关系。从该示范区高产奶牛筛选获得2头持续感染牛,分离获得1株非致细胞病变型BVDV,命名为NX2019/01。测序获得基因组全序列（12 107 nt）,其中ORF长11 703 nt,编码3 898个氨基酸。在基因组水平,NX2019/01株与我国SD-15、ZM-95、XC、LN-1等1m亚型分离株相似性较高（92.17%~93.84%）,但E^rns、E1以及E2基因存在较大差异。示范区同群牛急性感染BVDV时,毒株E2蛋白N端编码区核苷酸突变可导致第9位或第67位氨基酸变异。重组分析表明,NX2019/01株E2基因179—288位核苷酸区段以及ZM-95株E1基因168位—E2基因332位核苷酸区段存在相似的重组信号,可能由主要亲本SD-15株与次要亲本LN-1株重组形成,表明NX2019/01株、ZM-95株在演化进程中与SD-15株以及LN-1株或早期流行的高度相似毒株存在密切关联。本研究从持续感染高产奶牛分离获得了牛源BVDV-1m亚型毒株,在基因组水平厘清了BVDV-1m亚型毒株的进化关系,并首次发现同亚型BVDV毒株基因同源重组,为进一步研究BVDV在我国的演化规律奠定了基础。     

Seroepidemiologic survey for antibodies to selected viruses in the respiratory tract of lambs

A serologic study was conducted to determine the prevalence of antibodies to, and infection rate of, Mastadenovirus ovi 5, M ovi 6, parainfluenza-3 (PI-3) virus, bovine herpesvirus-1 (BHV-1), respiratory syncytial virus (RSV), bovine viral diarrhea (BVD) virus, and ovine progressive pneumonia (OPP) virus in lambs at a ram lamb growth-rate test station. For 2 consecutive years, serum samples were prepared from blood collected from 1- to 2-month-old ram lambs as they entered the test station (1st sample) and again 2 months later (2nd sample). The 1st year, 59 producers submitted 237 lambs; the 2nd year, 65 producers submitted 253 lambs. Microtitration serum virus-neutralization tests were used to determine antibody titers for M ovi 5, M ovi 6, PI-3 virus, BHV-1, and BVD virus. Antibodies to RSV and OPP virus were determined, using indirect hemagglutination and agar-gel immunodiffusion, respectively. Based on results of the 1st blood samples collected, the mean prevalence for both years was as follows: 95% of the lambs were seropositive for M ovi 5; 87.2% for PI-3 virus; 84.5% for RSV; 41.7% for M ovi 6; 8.7% for BVD virus; 5.4% for BHV-1; and 3.3% for OPP virus. Based on the 2-year mean, M ovi 6 had the highest infection rate (207 of 484 [42.8%]) as determined by the number of lambs evaluated having a greater than or equal to 4-fold increase in serum antibody titer from the 1st to the 2nd sampling. Infection rates of the other viruses were: 31.0% for M ovi 5; 15.3% for PI-3 virus; 5.6% for RSV; 0.6% for BVD virus; and 0.4% for BHV-1. One lamb became seropositive for OPP virus the 2nd year.     

Epidemiology and genetic characterization of BVDV,BHV-1, BHV-4, BHV-5 and Brucella spp. infections in cattle in Turkey

The aim of the study was to determine the epidemiological data of bovine viral diarrhea virus (BVDV), bovine herpesvirus-1 (BHV-1), bovine herpesvirus-4 (BHV-4), bovine herpesvirus-5 (BHV-5) and Brucella–associated cattle that were previously reported to have abortion and infertility problems in Ankara, Corum, Kirikkale and Yozgat provinces, Turkey. Whole blood and sera samples were obtained from 656 cattle, and antibodies against Brucella spp. were detected in 45 (6.86%) and 41 (6.25%) animals by Rose Bengal plate and serum tube agglutination tests, respectively. The seropositivity rates against BVDV, BHV-1 and BHV-4 were 70.89%, 41.3% and 28.78%, respectively. RT-PCR and PCR were performed to detect RNA and DNA viruses in blood samples, respectively. The BVDV 5′-untranslated region and BHV-1 gB gene detected in this study were phylogenetically analyzed. The BVDV strains analyzed in this study were closely related to those previously reported from Turkey. The nucleotide sequence from the BHV-1 strain detected in this study is the first nucleotide sequence of BHV-1 circulating in this area of Turkey deposited in the GenBank. The presence of Brucella spp. and prevalence of BHV-1, BHV-4 and BVDV in cattle should be further investigated throughout these regions.     

A long-term bacteriological and immunological study in Holstein-Friesian cattle experimentally infected with Mycobacterium avium subsp. paratuberculosis and necropsy culture results for Holstein-Friesian cattle, Merino sheep and Angora goats

The aims were to longitudinally evaluate the interferon-gamma (IFN-gamma) test in comparison to faecal culture and the absorbed ELISA in a cattle infection model for Johne's disease and to determine the adult infection status, by necropsy and tissue culture, of sheep, goats and cattle infected as young animals. Clinical disease, faecal culture results and immunological responses for Merino sheep [Stewart, D.J., Vaughan, J.A., Stiles, P.L., Noske, P.J., Tizard, M.L.V., Prowse, S.J., Michalski, W.P., Butler, K.L., Jones, S.L., 2004. A long-term study in Merino sheep experimentally infected with Mycobacterium avium subsp. paratuberculosis: clinical disease, faecal culture and immunological studies. Vet. Microbiol. 104, 165-178] and Angora goats [Stewart, D.J., Vaughan, J.A., Stiles, P.L., Noske, P.J., Tizard, M.L.V., Prowse, S.J., Michalski, W.P., Butler, K.L., Jones, S.L., 2006. A long-term study in Angora goats experimentally infected with Mycobacterium avium subsp. paratuberculosis: clinical disease, faecal culture and immunological studies. Vet. Microbiol. 113, 13-24], in the same experiments as the Holstein-Friesian cattle, have been described. Two longitudinal experiments involving Holstein-Friesian cattle challenged with either bovine or ovine strains of Mycobacterium avium subsp. paratuberculosis (Map) have been conducted over a period of 54 and 35 months, respectively. Blood samples for the IFN-gamma test and the absorbed ELISA and faecal samples for bacteriological culture were taken pre-challenge and monthly post-challenge. Cell-mediated (CMI) responses were substantially higher for the bovine Map strain during the 42-month period following dosing but then declined in the remaining 12 months. However, for the ovine Map challenge and control groups, CMI responses were not significantly different from each other. None of the cattle developed clinical disease and only one of the cattle in the bovine Map gut mucosal tissue challenged group was a persistent faecal shedder and also an ELISA antibody responder which developed after shedding commenced. Culture of tissues, following necropsy at the completion of the experiments, showed no evidence of infection in any of the challenged cattle and sheep for either the bovine or ovine Map strain in contrast to positive cultures for challenged goats in the same experiments. The tissues from the control cattle, sheep and goats were culture negative. The cattle were less susceptible to the bovine and ovine Map strains than goats and sheep with the goats being the least naturally resistant.     

Pestivirus infections in Norway. Serological investigations in cattle, sheep and pigs.

Serum samples from 1,133 dairy cows (187 herds), 3,712 ewes (103 flocks) and 1,317 adult pigs (877 herds), were tested for neutralizing antibodies against the NADL strain of bovine virus diarrhoea virus. The prevalence rate of seropositive animals was 18.5% in cattle, 4.5% in sheep and 2.2% in pigs, such seroreactors being found in 28% of the cattle herds and 18% of the sheep flocks. In all three species the rate showed considerable herd and geographical variation. In cattle the seroreactor rate was similar in herds with normal reproduction and in 62 herds with problems of repeat breeding. Of 31 pig sera containing antibodies against the NADL strain, 27 were also positive in a neutralization test for antibodies against swine fever virus (Baker strain). However, all sera showed a higher titre of antibodies against the bovine strain than against the swine fever virus. It was concluded that the immune response of the pigs had been induced by ruminant pestivirus, and not by swine fever virus.     

mcr-1 encoding colistin resistance in CTX-M-1/CTX-M-15- producing Escherichia coli isolates of bovine and caprine origins in Tunisia. First report of CTX-M-15-ST394/D E. coli from goats

The objective of this study was to isolate and characterize ESBL-producing Escherichia coli (ESBL-EC) from raw bovine and caprine milk samples, as well as from bovine faeces in Tunisia. Therefore, 120 bovine faecal samples and 9 caprine raw milk samples were collected from 2 extensive dairy-cow-farms and 5 ovine farms, respectively. In addition, 94 raw bovine milk samples, from containers and holding tanks from 50 small public-markets in the North of Tunisia, were processed for the isolation of cefotaxime-resistant E. coli (CTX^R). Antimicrobial susceptibility testing was carried out by disc-diffusion/broth-microdilution methods. The presence of genes encoding ESBL, as well as those encoding colistin (mcr-1 to 5 genes)- sulfonamide-, tetracycline-, gentamicin-, quinolone and chloramphenicol-resistance and class 1 integrons were tested by PCR (and sequencing in some cases). ESBL-EC isolates were further characterized by phylogrouping and MLST/PFGE typing. Eight samples (3.6%) contained ESBL-EC isolates (3/2 from raw bovine/goat milk and 3 from cattle faeces) and one isolate/sample was characterized. Four ESBL-EC isolates, all of bovine origin (3 faeces/1 milk), were resistant to colistin (MIC: 8–16 μg/ml), harboured the mcr-1 gene and carried IncP- and IncFIB-type plasmids. The 8 ESBL-EC strains had the following characteristics: a) bovine faeces: mcr-1/CTX-M-1/D-ST1642 (3 strains); b) raw milk: mcr-1/CTX-M-1/A-ST10 (1 strain); CTX-M-15/B1-ST394 (3 strains), and CTX-M-15/A-ST46 (1 strain). Most of bovine ESBL-EC isolates were multidrug-resistant (4/5). Our results showed that ESBL-EC were detected in bovine and caprine samples (CTX-M-1/CTX-M-15 producers), being some of them colistin-resistant (associated with mcr-1 gene), and they belonged to international clonal lineages.