Antimicrobial resistant Escherichia coli isolates in cattle and house sparrows on two Czech dairy farms

Rectal smears of calves, cows and young bulls, as well as cloacal smears of house sparrows (Passer domesticus), from farms at the villages of Sumice and Troskotovice, Czech Republic, were examined for E. coli resistant to 12 antimicrobials. The resistant isolates were tested for antimicrobial-resistance genes and integrons. Totals of 40% (n=183), 3% (n=95), 0% (n=33), and 9% (n=54) of Escherichia coli isolates from calves, cows, young bulls and house sparrows, respectively, were antimicrobial resistant. The following genes were identified in cattle E. coli isolates: tetA, tetB (isolates resistant to tetracycline), bla(TEM) (beta-lactams), strA, aadA (streptomycin), sul1, sul2 (sulphonamides), and cat, floR (chloramphenicol). Seven of 16 antimicrobial-resistant calf isolates from the Sumice farm possessed class 1 integrons with the aadA1 gene cassette integrated, 1 kb in size. On the Troskotovice farm, eight of 57 antimicrobial-resistant calf isolates possessed class 1 integrons. Integrons of 1.5kb with the dhfr1- aadA1 gene cassette were found in four isolates, followed by a 1kb integron with the aadA1 gene found in three isolates, and a 1.7kb integron with the dhfr17-aadA5 gene cassette and the phenotype ASSuTSxtNaCipCCfG. The prevalence of resistant E. coli in calves compared to adult cattle was much higher and probably was influenced by oral antimicrobial usage in calves, feeding with milk and colostrum from treated cows, as well as mechanisms unrelated to antimicrobial drug selection. Although house sparrows lived together with the cattle and came into contact with cattle waste on the farm, they were not infected by resistant E. coli isolates with the same characteristics as those found in cattle.     

Current understanding of the aetiology and laboratory diagnosis of footrot

Footrot is a highly contagious disease of the feet of ruminants caused by the synergistic action of certain bacterial species of which Dichelobacter nodosus (D. nodosus) is the main transmitting agent. The infection is specific to sheep and goats, although it has also been reported in cattle, horses, pigs, deer and mouflon. The antigenic diversity of D. nodosus is due to variations in the DNA sequence of its fimbrial subunit gene (fimA) and provides the basis for classification of the organism into at least 10 major serogroups (A-I and M), the distribution of which varies with different geographical locations. Host immune response to vaccination is serogroup specific. There are three different clinical forms of disease caused by virulent, intermediate and benign strains of D. nodosus, respectively. In order to facilitate rapid and reliable clinical diagnosis, virulence determination, strain differentiation and serogroup identification for effective control measures, immunological tests, DNA probes and PCR based techniques have been introduced. This review summarises the current understanding of the mechanisms of antigenic diversity of D. nodosus as well as advances made in its strain differentiation and diagnosis.     

Isolation and characterization of Dichelobacter nodosus from ovine and caprine footrot in Kashmir, India

Footrot is a highly contagious and economically important disease of sheep and goats, caused by Dichelobacter nodosus, a slow growing anaerobic Gram-negative rod. The current Australian antigenic classification system, based on variation in the fimbriae, classifies D. nodosus into at least 10 serogroups (A-I and M) and 18 serotypes. This investigation was intended to determine the serological diversity of D. nodosus in this region of Kashmir, India. Exudates of footrot lesions were collected from 24 naturally infected sheep and 42 goats located in the Kashmir valley. Of these 66 samples, 24 yielded evidence of D. nodosus by PCR using 16SrDNA specific primers. Multiplex PCR using serogroup specific primers revealed the presence of serogroup B in all the samples except two, which showed the presence of serogroup E D. nodosus. This study also documents the isolation of D. nodosus and detection of serogroup E for the first time in India.     

Occurrence of non-sorbitol fermenting, verocytotoxin-lacking Escherichia coli O157 on cattle farms

Escherichia coli O157 is often associated with hemorrhagic colitis and the hemolytic uremic syndrome (HUS). The verocytotoxins are considered to be the major virulence determinants. However, vt-negative E. coli O157 were recently isolated from patients with HUS. Several transmission routes to humans are described, but cattle feces are the primary source from which both the food supply and the environment become contaminated with E. coli O157.In a prevalence study performed on dairy, beef, mixed dairy/beef and veal farms in the summer of 2007, vt-negative isolates were detected on 11.8% (8/68) of the positive farms. From these eight farms, a total of 43 sorbitol-negative E. coli O157:H7 were collected. On five farms, only strains negative for the vt genes were present whereas both vt-negative and vt-positive strains could be detected on three other farms. Further characterization revealed that all isolates carried the eaeA and hlyA genes. Pulsed-field gel electrophoresis (PFGE) of all isolates resulted in nine different PFGE types and within the vt-negative strains, four different genotypes were identified, indicating that certain genetic clones are widespread over the cattle population.     

The in vitro cytotoxicity of native and modified trichloroacetic acid-extracted antigens from Fusobacterium necrophorum for mouse peritoneal macrophages

A Boivin lipopolysaccharide-protein complex (LPS-P), Westphal LPS (W-LPS), Boivin LPS (B-LPS), and a lipid A-associated protein (LAP) were extracted from Fusobacterium necrophorum bov 5 strain and chemically analyzed for protein, total nitrogen, hydrolyzed hexosamine, and nucleic acid content. Dose and time effects on the in vitro cytotoxicity of these preparations for mouse peritoneal macrophages were determined. Macrophage monolayers were exposed to 2.0 ml of 10, 100, 250, 500, or 1000 μg/ml solutions of the individual preparations for 1, 2, 4, or 6 hr at 37°C, and relative toxicities were determined; B-LPS> LPS-P> LAP, W-LPS. The effects of heat, NaOH, and trypsin and pronase on the native antigens were determined. Heat treatment had no effect on the cytotoxicity of B-LPS, but increased the toxicity of LPS-P and LAP for macrophages. Alkaline treatment decreased the cytotoxicity of B-LPS and increased the toxicity of LPS-P and LAP for macrophages. Enzyme treatment had no effect on LPS-P toxicity. The relationship of lipid A and protein content to toxicity for macrophages is discussed. Lipid A appears to be important in the cytotoxicity of B-LPS, and the protein components of LPS-P and LAP may contribute to their toxic activity. The ability of these bacterial cellular antigens to destroy phagocytic cells may facilitate the establishment of necrotic abscesses in susceptible hosts.     

Effect of biotin supplementation on hoof health and ceramide composition in dairy cattle

The effect of biotin supplementation on various foot lesions and hoof ceramide composition of toe (wall) and sole portions of hooves was studied in crossbred dairy cattle. Biotin supplementation was done for five months in 14 cattle at a farm and the other 14 animals kept as control. A significant decline was observed in heel erosions and sole avulsions along with total disappearance of white line fissures and double soles in the biotin supplemented cattle resulting in decrease in the overall disease score. Thin layer chromatographs of the hoof lipids revealed 11 types of ceramides in sole lipids and 6 types of ceramides in toe (wall) lipids. The ceramides were typed and identified according to their Rf values. A qualitative increase in the density of thin layer chromatographs of sole lipids was observed in biotin supplemented cattle whereas a non-significant difference in density of thin layer chromatographs of toe lipids was observed after supplementation of biotin.     

Prevalence of lameness and of associated claw disorders in Greek dairy cattle industry

The aim of this study was to assess the prevalence of lameness as well as the prevalence of claw–horn disruptions, abnormal claw shape and dermatitis in lame cows in Greek dairy farms and to evaluate their risk factors. Forty dairy farms were visited twice, during winter and during summer, and the lameness of milking cows was scored using a 5-point scale. In total 760 cows were lame (lameness score ≥ 3) and were further examined to identify macroscopically the claw disorders. The herd size, the trimming and footbathing frequency, the floor surface, the cleanness of the herd, the scraping frequency and the disinfectant used in the footbaths were recorded. The mean lameness prevalence was 18.7% and that of claw disorders observed in the lame cows was 75.4% for abnormal claw shape, 30.2% for dermatitis and 30.6% for claw–horn disruptions. Large herd size and the absence or only once per year trimming were associated with increased risk for the presence of lameness.     

Role of nitric oxide production in dairy cows naturally infected with Mycobacterium avium subsp. paratuberculosis

Nitric oxide (NO) is a crucial mediator in host defense and is one of the major killing mechanisms within macrophages. Its induction is highly affected by the types of cytokines and the infectious agents present. In the current study, NO production was evaluated after in vitro infection of unfractionated peripheral blood mononuclear cells (PBMCs) with Mycobacterium avium subsp. paratuberculosis (MAP) after 8 h, 3 and 6 days of culture for cows in different stages of disease. In addition, the effects of in vitro exposure to inhibitory cytokines such as interleukin-10 (IL-10) and transforming growth factor β (TGF-β) as well as the pro-inflammatory cytokine IFN-γ were correlated with the level of NO production. Nitric oxide production was consistently higher in cell cultures from subclinically infected animals at all time points. An upregulation of NO production was demonstrated in unfractionated cell cultures from healthy control cows after exposure to MAP infection as compared to noninfected cell cultures. A similar increase in NO due to the addition of MAP to cell cultures was also noted for clinically infected cows. NO level among subclinically infected cattle was greater at all time points tested and was further boosted with the combination of both in vitro MAP infection and IFN-γ stimulation. Alternatively, nonspecific stimulation with LPS from Escherichia coli O111:B4-W resulted in an upregulation of NO production in all infected groups at 3 and 6 days after in vitro infection. Finally, the in vitro exposure to inhibitory cytokines such as IL-10 and TGF-β prior to MAP infection or LPS stimulation resulted in the downregulation of this inflammatory mediator (NO) in all experimental groups at all time points. In summary, a higher level of NO production was associated with cows in the subclinical stage of MAP infection. As well, the results demonstrated an increase in NO production upon infection with MAP and in the presence of exogenous IFN-γ. Finally, the results suggest an important role of IL-10 and TGF-β on the profile of NO production which may explain the low NO production in MAP clinically infected cows.     

The impact of Theileria parva infections and other factors on calf mean daily weight gains in smallholder dairy farms in Murang’a District, Kenya

The association between mean daily weight gain, Theileria parva infections, clinical East Coast fever and other possible determinants of weight gain were examined in a longitudinal observational study that was conducted in cohorts of female calves from five agro-ecological zone (AEZ)-grazing strata. The strata were upper-midlands (UM) 1 zero-grazing, UM 1 open-grazing, UM 2 zero-grazing, UM 4 zero-grazing and UM 4 open-grazing. In total, 225 calves on 188 smallholder dairy farms were visited within the first 2 weeks of life and thereafter at biweekly intervals up to the age of 6 months between March 1995 and August 1996. During each visit, the calves were weighed and other calf-management practices in the farm during the visit such as housing, feeding and tick control also were recorded. Other events such as morbidity and mortality between or during the visits were also recorded.The overall mean daily weight gains were 0.24–0.29 kg (S.D.=0.17–0.22 kg) and were lower than the recommended targets for smallholder farms of 0.40–0.50 kg. The major tendency in variability of daily weight gains was due to visit-to-visit variation (especially in calves >3 months old).Differences in mean daily gains were associated with AEZ-grazing strata and calf-level factors that included breed of calf, calf sickness, incidence of ECF, feeding of milk, concentrate feeds and minerals and interaction between calf age and AEZ-grazing strata (P<0.05). ECF and other calf sicknesses exerted a temporal effect on calf-growth at the height of illness and immediately after; calves later recovered the lost growth except where other factors such as poor calf nutrition prevailed. Improvement in calf-growth in Murang’a District is achievable and extension services should continue to target individual-calf-level management practices.     

Antibody-ELISA for Trypanosoma evansi: Application in a serological survey of dairy cattle, Thailand, and validation of a locally produced antigen

Trypanosoma evansi is generally considered a mild pathogen in bovines. However, in Asia, acute and chronic signs have been observed in cattle, with high levels of parasitaemia, abortion and death. Investigations in Asian cattle are needed to better understand this epidemiological situation. To generate comparable data at a regional level, development and standardization of an antibody-enzyme linked immunosorbent assay for T. evansi (ELISA/T. evansi) was initiated and applied in an epidemiological survey carried out in dairy cattle in Thailand. A batch of 1979 samples was collected from dairy farms located throughout the country's four regions. Soluble T. evansi antigens initially produced in France were also produced in Thailand for comparison and technology transfer. Screening of 500 samples allowed us to identify reference samples and to determine the cut-off value of the ELISA. Seropositive animals – some of them confirmed by PCR – were found in the four regions, in 12 out of 13 provinces, in 22 out of 31 districts, in 56 farms out of 222 (25%, 95%CI ± 6%) and in 163 animals out of 1979 (8.2, 95%CI ± 1.2%). Estimated seroprevalence in 35 farms ranged between 1% and 30%, and in 21 farms it was >30%. Approximately 25% of survey cattle were exposed to the infection, in various situations. A sub-sample of 160 sera was tested on both antigens. Wilcoxon's (Z = 1.24; p = 0.22) and McNemars's tests (CHI2 = 3.55; p = 0.09) did not show any significant differences, showing that the locally produced antigen is suitable for further evaluation in the surrounding countries. Use of this standardized serological method will broaden knowledge of the prevalence and impact of the disease at the regional level in South-East Asia. Further validation of this ELISA will be necessary in other host species such as buffalo, horse and pig.     

The effects of liveweight loss and milk production on the risk of lameness in a seasonally calving,pasture fed dairy herd in New Zealand

Dairy herd managers have attempted to increase and maintain profits by selectively breeding dairy cattle for high production. Selection for milk production may have resulted in a tendency for greater liveweight (LW) loss postpartum. This study aimed to: (1) determine if excessive LW loss and milk yield in the first 50 days in milk (DIM) was associated with the development of lameness after 50 DIM, and (2) estimate the incidence risk of lameness in this herd attributable to excessive liveweight loss. The dataset comprised details from 564 mixed age cows from a single, seasonally calving, pasture fed dairy herd in New Zealand.     

Associations between management practices and colostrum quality on New Zealand dairy farms

Abstract

AIMS: To describe colostrum quality in spring-calving dairy herds in New Zealand, in terms of Brix, pH and total and coliform bacterial counts and to investigate associations between farm management practices and these measures of colostrum quality.

METHODS: In June 2015, commercial dairy farms (n=105), located in North and South Islands of New Zealand, were visited shortly after the first cows had calved, and when approximately 50% and 80% of the herd had calved (early, middle and late visits). One litre of pooled colostrum that was being fed to newborn calves was collected at each visit and used to determine Brix, pH, total bacterial and coliform counts. A survey of calf management practices was conducted with the herd manager or calf rearer after the final visit.

RESULTS: Of 298 pooled colostrum samples tested 29/298 (9.7%) had Brix >22%. Brix was higher on farms where calves were picked up twice daily compared with once daily (18.2 (95% CI=16.5–19.9)% vs. 15.9% (95% CI=15.2–16.6)%; p=0.012), and was lower where first milking colostrum was combined with colostrum obtained at later milkings (15.0 (95% CI=13.9–16.1)%) compared with where it was not (16.9 (95% CI=16.3–17.6)%; p=0.002). Vaccination of all cows against calf diarrhoeal pathogens was associated with increased Brix compared with no vaccination (18.1 (95% CI=16.6–19.6)% vs. 16.3 (95% CI=15.6–17.0)%; p=0.033). Mean pH of samples tested decreased from 5.97 (95% CI=5.84–6.09) to 5.58 (95% CI=5.45–5.71) for early and late-season visits, respectively (p<0.001). Of 268 samples tested, 23 (8.6%) had bacterial counts below the recommended threshold of 1.00×10⁵ cfu/mL. Mean bacterial counts increased from 2.75 (95% CI=1.80–3.70)×10⁸ to 4.99 (95% CI=3.95–6.03)×10⁸ cfu/mL for early and late-season visits, respectively (p<0.001). Of 259 samples tested, 23 (8.9%) had coliform counts below the recommended threshold of 1.00×10⁴ cfu/mL.

CONCLUSIONS AND CLINICAL RELEVANCE: On a large majority of dairy farms included in this study the pooled colostrum fed to newborn calves had sub-optimal Brix and excessive bacterial counts. Farm-level risk factors such as twice daily pick up of calves from the paddocks, herd vaccination and feeding pooled colostrum that did not combine colostrum obtained at later milkings with first milking colostrum were associated with colostrum quality measures.     

The first report of Histophilus somni pneumonia in Nigerian dairy cattle

Clinical signs of severe bronchopneumonia, including anorexia, coughing, nasal discharge, dyspnoea, diarrhoea, distension of the neck, lethargy, recumbency, lameness preceding collapse, and death were observed among a herd of Holstein–Friesian dairy cattle. The outbreak occurred over a 30-day period, and attack and case-fatality rates were 0.4% and 50%, respectively. At necropsy, extensive consolidation in the cranioventral parts of the lungs was observed. Histologically, a severe acute bronchopneumonia with slight pleuritis was present. Both pathological and bacteriological evaluation of the lungs incriminated Histophilus somni (heavy growth). Supplementary laboratory investigations also isolated Clostridium and Klebsiella species (scanty growth) from the lungs. Histophilosis in cattle was confirmed for the first time in Nigeria.     

Associations of Neospora caninum seropositivity with gestation number and pregnancy outcome in Danish dairy herds

The prevalence and distribution of seropositivity towards the protozoan parasite Neospora caninum were studied in single blood samples from 1561 cows from 31 Danish dairy herds. Blood samples were analysed by an indirect enzyme-linked immunoassay and an indirect fluorescent-antibody test. Seroprevalence in 15 herds with previous abortions assigned to neosporosis ranged from 1% to 58%, with a mean frequency of 22%. In eight out of 16 herds without a history of N. caninum related abortions, no seroreactors were found. In the remaining eight herds, the seroprevalence ranged from 6% to 59%. The prevalence and distribution of seropositivity, gestation number prior to sampling, and breed were related to abortions and perinatal deaths using a random-effects logistic-regression model. Abortion risk was significantly increased in seropositive animals (OR=3) and in 2nd-gestation cows (OR=3). Perinatal death was significantly influenced by gestation number and breed, but not by serostatus. Reproductive performance and culling risk of cows were not affected by serostatus. Seropositivity increased with “age” (i.e. gestation number) (P=0.02). In open cows, seropositivity tended to decrease with distance from calving (P=0.05). The proportion of seropositive pregnant cows increased with trimester (P=0.02).     

Difficulties to control natural infestation with Otobius megnini (Acari: Argasidae) nymphs in cattle with systemic biocides

A study was carried out to determine the efficacy of topical eprinomectin against nymphal infestation of Otobius megnini in cattle, where a group of 14 individuals were treated with a dose of 0.5 mg/per kg of body weight applied on the dorsal midline, and a group of 14 individuals remained as control. Tick burdens between treated and control groups showed no statistically significant differences (P > 0.05), and the mortality of the nymphs was similar in both groups (P: 0.828). All females obtained from nymphs of both groups were able to copulate with males from the corresponding group and laid eggs that produced visually normal larvae. The failure of eprinomectin and other biocides applied per os or by injection to control nymphs of O. megnini in cattle indicate that sanitary measures and applications of biocides into the ears would represent the most reasonable approach to control of this tick on cattle.     

Prevalence and molecular characterization of bovine Cryptosporidium in Qazvin province, Iran

The aim of this study was to determine the prevalence, variability with host age, and the genotypes of species of Cryptosporidium in cattle from 15 dairy farms in Qazvin province, Iran. Fecal samples, collected from 272 cattle during May 2006 to December 2007, were characterized microscopically. Oocysts from 51 positive samples were analyzed using PCR assay of 18S SSU rRNA, restriction fragment length polymorphism (RFLP) and sequencing. We identified 72.6% of the positive samples as Cryptosporidium parvum, 17.7% as Cryptosporidium andersoni, 7.8% as Cryptosporidium bovis and 1.9% as a novel genotype of C. parvum possessing a single mutation on MboII restriction. An infection rate of 19.5% of C. parvum among 174 pre-weaned calves was significantly higher than the 3.1% among 98 post-weaned calves (P < 0.0006). This is the first report of C. bovis and the new subgenotype of C. parvum in Iranian cattle.     

Prevalence of species and genotypes of Cryptosporidium found in 1-2-year-old dairy cattle in the eastern United States

The prevalence of Cryptosporidium species in 1-2-year-old heifers was determined for 571 animals on 14 dairy farms in seven states on the East Coast of the United States. A fecal specimen collected directly from each heifer was processed to concentrate oocysts that were then examined by polymerase chain reaction (PCR). For every PCR-positive specimen the 18S rRNA gene of Cryptosporidium was sequenced. Cryptosporidium was identified by PCR from heifers on 13 of 14 farms. On all except four farms groups of heifers were housed in a barn or in large covered pens. Others were pastured. From many of the same farms an earlier study reported that 41% of 393 pre-weaned calves and 26.2% of 447 post-weaned calves were infected. In the present study, 11.9% of 571 heifers were infected with Cryptosporidium, 0.7% with Cryptosporidium parvum, the zoonotic species. Of 68 PCR-positive specimens characterized by gene sequencing 1, 4, 10, 24, and 29 calves were infected with Cryptosporidium suis, Cryptosporidium parvum, Cryptosporidium deer-like genotype, Cryptosporidium bovis, and Cryptosporidium andersoni, respectively. These findings demonstrate a lower prevalence of infection in 1-2-year-old dairy cattle than in younger cattle as well as a change in the diversity of species present. Consequently, the risk of humans acquiring infection with C. parvum from exposure to feces from yearling and older cattle appears much lower than from exposure to pre-weaned calves.     

The effect of hindclaw height differential and subsequent trimming on lameness in large dairy cattle herds in Canterbury,New Zealand

Abstract

AIMS: To determine if inequality in height between claws within each hindlimb of dairy cattle is a risk factor for the development of lameness and to determine the effect of preventive trimming of these claws on the subsequent risk of lameness.

METHODS: Cows (n=2,695) on three dairy farms in Canterbury, New Zealand, were examined and the height differential between their hind claws on each limb was measured. Cows were blocked by age, then breed and grouped by height differential, before being randomly assigned to either treatment or control groups. Cows in the control group were not trimmed, whilst cows in the treatment group were all trimmed according to the ‘Dutch Trimming’ method by an experienced foot trimmer. All subsequent clinical incidences of lameness were identified, recorded and classified according to five categories. The overall incidence of lameness, the effect of claw height differential on the incidence of lameness and the effect of trimming on the incidence of lameness were assessed using both GEE modelling and survival analysis.

RESULTS: The incidence of lameness identified in any untrimmed hindlimb irrespective of diagnosis, measured at a limb level and assessing the first incidence of lameness only, was 10.0% for a 6-month period from November to drying off. The predominant lesion identified was white line disease (WLD; 7.9% incidence and 78.6% of all lameness). The overall incidence of hindlimb lameness measured at cow level for the same period was 14.5%. The risk of lameness was influenced by breed (p=0.016), age (p=0.002) and claw height differential (p=0.026). There were no significant interactions. The risk of lameness was lowest where the claw height differential was 0–2 mm. The incidence of lameness in the first 70 days following trimming was higher in older cows than in younger cows (p<0.001). Trimming did not affect lameness incidence (p=0.185). The Cox proportional hazard model demonstrated that farm (p<0.001), trimming group (p=0.021); and age (p=0.021) significantly affected the median days to lameness in the first 70 days following trimming.

CONCLUSIONS AND CLINICAL RELEVANCE: Hind feet with lower height differentials between claws had a lower incidence of lameness. Trimming of these feet to remove the height differential did not reduce their overall incidence of lameness, but did increase days to first lameness.     

Seroprevalence of Toxoplasma gondii and Neospora caninum in feral cats (Felis silvestris catus) in Majorca, Balearic Islands, Spain

Cryptosporidium spp. are common intestinal protozoan parasites that infect a wide range of hosts, including humans and livestock, worldwide. The objective of this study was to determine the prevalence of Cryptosporidium spp. in dairy calves in Prince Edward Island, Canada, and the potential for transmission of this parasite between dairy calves and humans. Fecal samples were collected from 183 dairy calves from 11 farms in Prince Edward Island. The prevalence of Cryptosporidium spp. infections in these animals was determined by examining for the presence of oocysts in the fecal samples, using immunofluorescence microscopy. Molecular characterization was done using a nested-PCR protocol to amplify fragments of the Cryptosporidium heat-shock protein 70 gene, followed by DNA sequencing. Ten calves (6.2%), representing 4 out of 11 farms tested, were positive for Cryptosporidium spp. DNA sequence analysis on five PCR positive samples demonstrated that Cryptosporidium parvum was the only species present in the calves tested, suggesting that there is a potential risk of zoonotic transmission between dairy calves and humans in this region.