Feces Of Calves Research Articles

A Comparison of Eggs Per Gram (Epg) from Wet Faecal Smears Standard Method, with Benchtop Flotation with Centrifugal Techniques in Determining the Number of GIN Eggs in Calf Faeces-A Case Study

Aim: Although several techniques exist for quantifying GastroIntestinal Nematode eggs (GIN), the method producing the highest egg counts would result in better targeting of parasitic control in livestock. Materials and Techniques: An experiment was therefore, conducted to determine the best technique in quantifying Gastrointestinal (GIN) helminths eggs in calf faeces by comparing the wet faecal smear, faecal floatation and centrifugation floatation techniques offaeces of a moderately parasitized calfinfested withan initial egg per gram (epg) of nine hundred and fifty (950). Seven (7) techniques were used and observations recorded in triplicate by separate observers.The solutions used for the quantification of were sodium chloride (NaCl), zinc sulphate (ZnSO4) and sucrose. The average eggs count from six techniques were tested and findings using descriptive statistics and an ANOVA due to technique are reported. Results: Descriptive statistics showed that centrifugation floatation techniques showed the highest quantification of GIN eggs. Egg counts by centrifugation were highest using floatation solutions sodium chloride followed by sucrose then zinc sulphate. These findings corresponded with high the significant difference (P Conclusion: Therefore, we recommend that centrifugation techniques be used for quantifying GIN eggs in targeting follow up anthelmintic treatment in livestock affected by gastrointestinal parasites. Keywords: GIN eggs, Solus, Eggs per gram, Faeces, Centrifugation, Floatation

Antimicrobial resistance status of Enterococcus from Australian cattle populations at slaughter.

Antimicrobial agents are used in cattle production systems for the prevention and control of bacterial associated diseases. A consequence of their use is the potential development of antimicrobial resistance (AMR). Enterococcus faecium and Enterococcus faecalis that are resistant to antimicrobials are of increased concern to public health officials throughout the world as they may compromise the ability of various treatment regimens to control disease and infection in human medicine. Australia is a major exporter of beef; however it does not have an ongoing surveillance system for AMR in cattle or foods derived from these animals. This study examined 910 beef cattle, 290 dairy cattle and 300 veal calf faecal samples collected at slaughter for the presence of enterococci. Enterococcus were isolated from 805 (88.5%) beef cattle faeces, 244 (84.1%) dairy cattle faeces and 247 (82.3%) veal calf faeces with a total of 800 enterococci subsequently selected for AMR testing. The results of AMR testing identified high levels of resistance to antimicrobials that are not critically or highly important to human medicine with resistance to flavomycin (80.2%) and lincomycin (85.4–94.2%) routinely observed. Conversely, resistance to antibiotics considered critically or highly important to human medicine such as tigecycline, daptomycin, vancomycin and linezolid was not present in this study. There is minimal evidence that Australian cattle production practices are responsible for disproportionate contributions to AMR development and in general resistance to antimicrobials of critical and high importance in human medicine was low regardless of the isolate source. The low level of antimicrobial resistance in Enterococcus from Australian cattle is likely to result from comprehensive controls around the use of antimicrobials in food-production animals in Australia. Nevertheless, continued monitoring of the effects of all antimicrobial use is required to support Australia’s reputation as a supplier of safe and healthy food.

Feeding of waste milk to Holstein calves affects antimicrobial resistance of Escherichia coli and Pasteurella multocida isolated from fecal and nasal swabs

The use of milk containing antimicrobial residues in calf feeding programs has been shown to select for resistant fecal Escherichia coli in dairy calves. However, information is scarce about the effects of feeding calves waste milk (WM) on the prevalence of multidrug-resistant bacteria. The objective of this study was to determine the antimicrobial resistance patterns of fecal E. coli and nasal Pasteurella multocida isolates from calves fed either milk replacer (MR) or WM in 8 commercial dairy farms (4 farms per feeding program). Fecal and nasal swabs were collected from 20 ± 5 dairy calves at 42 ± 3.2 d of age, and from 10 of these at approximately 1 yr of age in each study farm to isolate the targeted bacteria. Furthermore, resistance of E. coli isolates from calf-environment and from 5 calves at birth and their dams was also evaluated in each study farm. Resistances were tested against the following antimicrobial agents: amoxicillin-clavulanic acid, ceftiofur, colistin, doxycycline (DO), enrofloxacin (ENR), erythromycin, florfenicol, imipenem, and streptomycin. A greater number of fecal E. coli resistant to ENR, florfenicol, and streptomycin and more multidrug-resistant E. coli phenotypes were isolated in feces of calves fed WM than in those fed MR. However, the prevalence of fecal-resistant E. coli was also influenced by calf age, as it increased from birth to 6 wk of age for ENR and DO and decreased from 6 wk to 1 yr of age for DO regardless of the feeding program. From nasal samples, an increase in the prevalence of colistin-resistant P. multocida was observed in calves fed WM compared with those fed MR. The resistance patterns of E. coli isolates from calves and their dams tended to differ, whereas similar resistance profiles among E. coli isolates from farm environment and calves were observed. The findings of this study suggest that feeding calves WM fosters the presence of resistant bacteria in the lower gut and respiratory tracts of dairy calves.

Using Duddingtonia flagrans in calves under an organic milk farm production system in the Mexican tropics

The reduction of the gastrointestinal nematode (GIN) larvae population in faeces of cattle treated with Duddingtonia flagrans chlamydospores on a farm under an organic production system in Chiapas, Mexico, was assessed. Seventeen Cebu/Swiss crossbreed grazing calves naturally infected with GIN, were randomly distributed into two groups and treated as follows: Group 1, an oral administration of 2 × 106D. flagrans chlamydospores/kg BW, every two days for 30 days; group 2, Control, without any treatment. Results indicated that the epg values in both groups remained similar (p > 0.05). The average number of (L3) from coprocultures from the group treated with D. flagrans had an important reduction (53.8%) with respect to the control group and it reached 75.3% maximum larval reduction at the 14th sampling; although, no statistic significance was observed (p > 0.05). Likewise, the average of larvae (L3) recovered from grass corresponding to the animals treated with D. flagrans diminished at 25.1% with respect to the control group (p > 0.05). A mixture of GIN genera including Strongyloides sp., Haemonchus sp., Cooperia sp., Trichostrongylus sp., Oesophagostomum sp. and Mecistocirrus sp., were identified from coprocultures. It was concluded that treatment with D. flagrans chlamydospores reduces the GIN larvae population in grass and in faeces of calves maintained under an organic milk production system.

自然哺乳子ウシから分離されたLactobacillus amylovorus Strain Kumamoto (JCM14931)の整腸剤としての可能性

自然哺乳子ウシから分離されたLactobacillus amylovorus Strain Kumamoto (JCM14931)の整腸剤としての可能性

Health, physiology, and behavior of dairy calves reared on 4 different substrates

The objective of this study was to compare the health, physiology, and behavior of group-housed calves reared on wood shavings with those reared on alternative surfaces. At 1 wk of age, 80 calves were moved into 1 of 20 experimental pens (n = 4 calves/pen) where they remained until 6 wk of age. Pens had floors covered with pea gravel (PG), rubber chip (RC), sand (SA), or wood shavings (WS; n = 5 pens/substrate). Body weight, cleanliness, health, and skin surface and vaginal temperature were recorded at 1, 3, and 6 wk of age. Escherichia coli numbers were assessed on the skin surface of the shoulder and in the feces of calves at 3 and 6 wk of age. Blood samples were taken at 1, 3, and 6 wk of age to measure hematological values and cortisol, IgG, and lactate concentrations. Behaviors (lying, running, and self-grooming) were recorded in the home pen at 1, 3, and 6 wk of age using video recorders and accelerometer data loggers. At 6 wk of age, calves were tested individually in an arena test and behavior was recorded continuously for 20 min. Body weight did not differ among calves reared on PG, RC, SA, or WS, regardless of age. All calves were clean and no calves displayed any signs of lameness, leg lesions, or injuries at wk 1, 3, or 6, regardless of substrate. The number of E. coli recovered from a surface area of 100 cm2 on the shoulder of each calf was affected by rearing substrate, with more E. coli recovered from calves reared on WS than PG, RC, or SA at 3 and 6 wk of age. Fecal E. coli counts were not affected by rearing substrate at 3 or 6 wk of age. Over the entire study period, calves reared on PG and SA had lower skin temperatures than calves reared on RC or WS, but skin temperature was similar between calves reared on PG and SA. However, vaginal temperature did not differ among calves reared on different substrates at 1, 3, or 6 wk of age. Hematology values and cortisol, IgG, and lactate concentrations of calves were similar among rearing substrates over the 6-wk study period. In the home pen, rearing substrate did not influence time spent lying; however, calves reared on WS performed more lying bouts than calves reared on PG or SA. In addition, rearing substrate did not influence the time calves spent running; however, calves reared on WS spent more time self-grooming than calves reared on PG, RC, and SA. During a 20-min arena test, running, bucks, jumps, and kicks performed by calves was not affected by rearing substrate. In conclusion, the physiology and behavior of calves reared on PG, RC, and SA was similar to WS, which is considered the preferred rearing substrate to use when rearing calves. Therefore, PG, RC, and SA may be acceptable substrate options when rearing group-housed dairy calves.

Identification of a novel bovine enterovirus possessing highly divergent amino acid sequences in capsid protein

BackgroundBovine enterovirus (BEV) belongs to the species Enterovirus E or F, genus Enterovirus and family Picornaviridae. Although numerous studies have identified BEVs in the feces of cattle with diarrhea, the pathogenicity of BEVs remains unclear. Previously, we reported the detection of novel kobu-like virus in calf feces, by metagenomics analysis. In the present study, we identified a novel BEV in diarrheal feces collected for that survey. Complete genome sequences were determined by deep sequencing in feces. Secondary RNA structure analysis of the 5′ untranslated region (UTR), phylogenetic tree construction and pairwise identity analysis were conducted.ResultsThe complete genome sequences of BEV were genetically distant from other EVs and the VP1 coding region contained novel and unique amino acid sequences. We named this strain as BEV AN12/Bos taurus/JPN/2014 (referred to as BEV-AN12). According to genome analysis, the genome length of this virus is 7414 nucleotides excluding the poly (A) tail and its genome consists of a 5′UTR, open reading frame encoding a single polyprotein, and 3′UTR. The results of secondary RNA structure analysis showed that in the 5′UTR, BEV-AN12 had an additional clover leaf structure and small stem loop structure, similarly to other BEVs. In pairwise identity analysis, BEV-AN12 showed high amino acid (aa) identities to Enterovirus F in the polyprotein, P2 and P3 regions (aa identity ≥82.4%). Therefore, BEV-AN12 is closely related to Enterovirus F. However, aa sequences in the capsid protein regions, particularly the VP1 encoding region, showed significantly low aa identity to other viruses in genus Enterovirus (VP1 aa identity ≤58.6%). In addition, BEV-AN12 branched separately from Enterovirus E and F in phylogenetic trees based on the aa sequences of P1 and VP1, although it clustered with Enterovirus F in trees based on sequences in the P2 and P3 genome region.ConclusionsWe identified novel BEV possessing highly divergent aa sequences in the VP1 coding region in Japan. According to species definition, we proposed naming this strain as “Enterovirus K”, which is a novel species within genus Enterovirus. Further genomic studies are needed to understand the pathogenicity of BEVs.

Composition of the microbiota in forestomach fluids and feces of Japanese Black calves with white scours.

The objective of this study was to characterize the composition of the forestomach and fecal microbiota in Japanese Black calves with white scours. Forestomach fluid, feces, and peripheral blood were collected from healthy calves ( = 5; age 10 ± 2 d) and scouring calves ( = 5; age 10 ± 1 d) on the day on which white scours occurred. The pH and concentrations of VFA, lactic acid, and ammonia nitrogen (NH-N) of the forestomach fluids were determined. Microbiota composition and gene copy numbers in the forestomach fluid and feces were analyzed by 454 pyrosequencing and quantitative real-time PCR (qPCR), respectively. The cytokine mRNA level in peripheral leukocytes was evaluated by qPCR. The pH of the forestomach fluid of the scouring calves tended to be higher than that of the healthy calves ( = 0.056). No significant difference was detected in the total VFA, lactic acid, or NH-N concentrations in the forestomach fluids of the 2 groups. Firmicutes, Bacteroidetes, and Proteobacteria were the predominant phyla in the forestomach fluid and feces. At the genus level, the relative abundance of in the forestomach fluid was significantly higher in the scouring calves ( < 0.05) and the relative abundance of in the feces was significantly higher than that in the forestomach in the healthy calves ( < 0.05). Furthermore, the bacterial diversity indices of feces were lower in the scouring calves. Quantitative PCR amplification using some of the primer pairs failed in the forestomach fluid and feces in both groups. These results suggested that fermentation in the forestomach may affect the occurrence of white scours, resulting in changes in the composition and diversity of the forestomach fluid and fecal microbiota in Japanese Black calves.

Occurrence of faecal shedding of Mycobacterium avium subs. paratuberculosis by calves

Paratuberculosis or Johne´s disease is a chronic granulomatous enteric disease afflicting cattle, sheep, goats and other ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Calves are infected mainly via an oral route, especially from manure and environment contamination by infected adult cattle. It is generally assumed that calves rarely shed MAP bacteria and that calf-to-calf transmission is of minor importance. The aim of the study was to evaluate occurrence of faecal excretion of MAP by calves born to MAP-positive and MAP-negative dams in a low MAP prevalence herd. The study involved 66 dam-calf pairs. Samples of faeces were collected: for each cow once about 2–3 weeks to parturition, and for each calf 3 times at 2–5, 180–185 and 360–365 days of age. All samples were studied for the presence of MAP by culture or by PCR of IS900 insertion sequence MAP DNA. Faecal samples of the dams demonstrated 15/66 dams with at least one positive result. Analysis of MAP shedding prevalence in the faeces of calves originating from MAP-positive and MAP-negative dams shows that in 15 MAP-positive dams- 10 out of 15 calves (66.66%) at the age of 180–185 days and at the aged of 360–365 days 11 out of 15 calves (73.33%) were MAP-positive in faecal samples. In the case of MAP-negative dams: 7 out of 51 calves (13.72%) at the age of 180–185 days, and 8 out of 51 calves (15.68%) at the age of 360–365 days were MAP-positive. MAP-infection cases in calves coming from infected and healthy dams prove the occurrence of calf-to-calf transmission and that contact with infected calves increases the risk of MAP spreading among healthy calves.

Antimicrobial activity of lactic acid bacteria multiplied in an alternative substrate and their influence on physiological parameters of new-born calves

Here, the ability of Pediococcus pentosaceus and Pediococcus acidilactici to utilise potato tuber juice for cell synthesis without an external nutrient supplement was investigated, and the influence of lactic acid bacteria (LAB) grown in this substrate on the growth performance of new-born calves, as well as blood biochemical and faecal microbiological parameters was evaluated. Calves were selected based on the analogy principle, treatment group (n = 21), control group (n = 27). Calves in the treatment group were administered 50 ml of fermented potato tubers juice containing 9.6 log CFU/ml of LAB mixture for 14 days. Also, determination of antimicrobial activities of tested LAB against a variety of pathogenic and opportunistic bacterial strains previously isolated from diseased cattle was performed. It was found that LAB supernatants effectively inhibited the growth of Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Salmonella enterica, Corynebacter spp., Klebsiella pneomoniae, Enterococcus faecalis, and Bacillus cereus (the diameters of the inhibition zones varied between 11.0 ± 0.3 mm and 17.0 ± 0.6 mm). Thus, potato juice can be used as an alternative substrate for LAB cultivation (LAB cell concentration 9.6 ± 0.07 log CFU/ml). After lyophilisation (-48 °C) and spray-drying (+150 °C) viable cell concentrations in the fermented potato juice powder were 9.18 ± 0.09 log CFU/g and 9.04 ± 0.07 log CFU/g, respectively. The 50 ml of fermented potato tuber juice containing 9.6 log CFU/ml of LAB, administered every day for 14 days, reduced the risk of developing acidosis (stabilised blood pH; P < 0.05), reduced lactates and PCO2 concentration (P < 0.05) and the risk of liver lesions (reduced serum alanine aminotransferase concentration; P < 0.005) in blood and E. coli in the faeces of new-born calves.

Characterization of virulence factors and phylogenetic group determination of Escherichia coli isolated from diarrheic and non-diarrheic calves from Brazil

This study aimed to detect virulence factors, pathovars, and phylogenetic groups of Escherichia coli strains obtained from feces of calves with and without diarrhea up to 70days old and to determine the association between occurrence of diarrhea, phylogenetic groups, and pathovars. Phylo-typing analysis of the 336 E. coli strains isolated from calves with Clermont method showed that 21 (6.25%) belong to phylogroup A, 228 (67.85%) to phylogroup B1, 2 (0.6%) to phylogroup B2, 5 (1.49%) to phylogroup C, 57 (16.96%) to phylogroup E, and 3 (0.9%) to phylogroup F. Phylogroup D was not identified and 20 strains (5.95%) were assigned as "unknown." The distribution of phylogenetic groups among pathovars showed that NTEC belong to phylogroups B1 (17) and C (4); EPEC to phylogroups B1 (6) and E (8); STEC to phylogroups A (5), B1 (56), B2 (2), C (1), and E (15); EHEC to phylogroups B1 (95) and E (5); and ETEC to phylogroups A (3), B1 (7), and E (10). The EAST-1 strains were phylogroups A (13), B1 (47), E (19), and F (3); E. coli strains of "unknown" phylogroups belonged to pathovars EPEC (1), EHEC (2), STEC (7), and EAST-1 strains (6). ETEC was associated with diarrhea (P=0.002). Our study did not find association between the phylogenetic background and occurrence of diarrhea (P=0.164) but did find some relationship in phylogenetic group and pathovar. The study showed that EHEC and STEC are classified as phylogroup B1, EAST-1 phylogroup A, ETEC, and EPEC phylogroup E.

Occurrence, genotyping, shiga toxin genes and associated risk factors of E. coli isolated from dairy farms, handlers and milk consumers

The objectives of the current study were to determine the occurrence and genotypes of E. coli in dairy farms, workers and milk consumers and to evaluate risk factors associated with contamination of milk in dairy farms. Molecular characterization of shiga toxin associated genes and enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) finger printing of E. coli from different sources were also studied. Paired milk samples and rectal swabs from 125 dairy cows, rectal swabs from 82 calves and hand swabs from 45 dairy workers from five dairy farms were collected. In addition, 100 stool samples from 70 diarrheic and 30 healthy humans were collected and examined for the presence of E. coli.E. coli was isolated from milk (22.4%), dairy cattle feces (33.6%), calf feces (35.4%), dairy worker hand swabs (11.1%) and stools of milk consumers (2%, from diarrheic patients only). Only stx1 was identified in seven of 12 E. coli O125 isolated from different sources. High genetic diversity was determined (Simpson's index of diversity, D = 1) and E. coli O125 isolates were classified into 12 distinct profiles, E1–E12. The dendrogram analysis showed that two main clusters were generated. Mastitis in dairy cows was considered a risk factor associated with contamination of the produced milk with E. coli. The isolation of E. coli from rectal swabs of dairy cows and calves poses a zoonotic risk through consumption of unpasteurized contaminated dairy milk. Educational awareness should be developed to address risks related to consumption of raw milk.

Longitudinal study of CTX-M ESBL-producing E. coli strains on a UK dairy farm

The aim of this study was to investigate the bacterial strains and farm environment that may contribute to the persistence of ESBL-producing E. coli on a single UK dairy farm. A longitudinal study was conducted comprising 6 visits, between August and October 2010, followed by a further visit at approximately 69weeks after the initial visit. Faecal and environmental samples were collected from different parts of the farm. The persistence and extent of faecal shedding of ESBL E. coli by individual calves was also determined. Twenty two different PFGE types were identified. Four of these were persistent during the study period and were associated with serotypes: O98, O55, O141 and O33. The counts suggest that shedding in calf faeces was an important factor for the persistence of strains, and the data will be useful for parameterising mathematical models of the spread and persistence of ESBL strains within a dairy farm.

Multilocus typing of Cryptosporidium spp. in young calves with diarrhea in Korea

We assessed the prevalence and performed molecular analysis of Cryptosporidium spp. in diarrheal feces of calves in Korea. Diarrheal fecal samples were collected from 951 young calves (<3months) on 425 farms. Cryptosporidium prevalence was assessed by PCR and ELISA, and molecular characterization was performed by targeting the 18S rRNA, heat-shock protein 70 (hsp70), and glycoprotein 60 (gp60) genes. Data were analyzed according to the sex, type of cattle, region, season, and type of diarrhea. PCR analysis revealed Cryptosporidium spp. in 9.9% (94/951) of diarrheal fecal samples. C. parvum and C. bovis/ryanae were present in 6.1% (58/951) and 4.1% (39/951) of diarrheal fecal samples, respectively. In addition, ELISA showed positive results for C. parvum in 9.7% (92/951) samples. Statistical analysis of the PCR and ELISA results revealed a lower prevalence of C. parvum in the hemorrhagic diarrheal samples (P<0.05). For C. bovis/ryanae, seasonality and high prevalence in hemorrhagic diarrhea were observed (P<0.05). Of the 951 samples tested for C. parvum, 903 samples showed agreement with a κ value of 0.65, indicating good agreement between the two tests. Although C. bovis and C. ryanae share highly similar 18S rRNA sequences, PCR based on hsp70 successfully distinguished C. bovis from C. ryanae. Sequence analysis of gp60 revealed that C. parvum belonged to the IIa families and was further subtyped as IIaA18G3R1 and IIaA16G3R1, which have not been previously reported in Asia. These findings indicate that Cryptosporidium spp. play an important role in diarrhea in young calves in Korea. Considering the zoonotic significance of C. parvum IIa subtype and dense rearing system of cattle in Korea, prevention and continuous monitoring of Cryptosporidium are required.

Genomic analysis of three Bifidobacterium species isolated from the calf gastrointestinal tract.

Ruminant animals contribute significantly to the global value of agriculture and rely on a complex microbial community for efficient digestion. However, little is known of how this microbial-host relationship develops and is maintained. To begin to address this, we have determined the ability of three Bifidobacterium species isolated from the faeces of newborn calves to grow on carbohydrates typical of a newborn ruminant diet. Genome sequences have been determined for these bacteria with analysis of the genomes providing insights into the host association and identification of several genes that may mediate interactions with the ruminant gastrointestinal tract. The present study provides a starting point from which we can define the role of potential beneficial microbes in the nutrition of young ruminants and begin to influence the interactions between the microbiota and the host. The differences observed in genomic content hint at niche partitioning among the bifidobacterial species analysed and the different strategies they employ to successfully adapt to this habitat.

Prevalence of Escherichia coli adhesion-related genes in neonatal calf diarrhea in Uruguay.

Neonatal calf diarrhea (NCD), one of the most important diseases of neonatal dairy and beef calves in Uruguay, has become relevant in association with intensive systems. This disease generates substantial economic losses every year worldwide as a result of increased morbidity and mortality. Escherichia coli, one of the pathogens associated with NCD, can express several fimbrial and afimbrial adhesins. The objective of this study was to assess the presence of clpG, f5, f17A, f17G(II), and f17G(I) genes that encode three important adhesins expressed in diarrheagenic E. coli: F5, F17 and CS31A, isolated from feces of calves in Uruguay. Feces of 86 (70 diarrheic and 16 healthy) calves, from 15 animal facilities in Uruguay, were collected between 2012 and 2013. Biochemical and molecular identification were performed to finally obtain 298 E. coli isolates. Partial amplification of adhesion-related genes was performed by polymerase chain reaction. The most prevalent gene was f17A (31.2%), followed by f17G(II), clpG, f17G(I) and f5 (25.8%, 17.5%, 3.7% and 0.7%, respectively). All genes were present in diarrheic and healthy animals except f5 and f17G(I); these genes were present only in affected calves, although in low numbers. This is the first report of the presence of F5, F17, and CS31A genes in E. coli strains from NCD cases in Uruguay. Prevalence values of the genes, except f5, were in accordance with regional findings. It is expected that further characterization of locally transmitted strains will contribute to control a problem of regional and international magnitude.

Bovine Coronavirus (BoCV) Infection in Calves with Diarrhoea and Their Dams

Background: Bovine coronavirus (BoCV) is common with high seroprevalence in dairy cattle. It is reported in many countries. Also, BoCV causes diarrhea in dairy calves. The transmission of BoCV is the fecal-oral/aerosol-nasal routes. Feces from clinical cases or clinically normal dairy cattle are source of infection, also contamination of feed and water. The purpose of the current study was to investigate BoCV infection in diarrheic calves (age and sex) and their dams (age). For this reason, the serological and virological methods were used. Haematological parameters of the calves and their dams were compared using the statistical methods.Materials, Methods &amp; Results: In this study, following clinical examination of 3500 cattle and their calves from 25 number of dairy farms 184 calves with diarrhoea and their dams (183) (≥2 - ≤6 age) were sampled for BoCV presence by enzyme linked immunosorbent assay (ELISA). Additionally, all blood samples were examined by hematological methods. 172 (93.99%) cows and 172 (93.99%) calves were found antibodies (Ab) positive (+). The high levels of Ab for BoCV were detected as 36.05 % in dams 6 years and older ages. In the calves, Ab to BoCV were found at the highest level (25.26%) in the female calves ≥5 - ≤6 months ages. BoCV antigen (Ag) was detected in only faecal sample of a (0.54%) calf. When the haematological parameters were compared between BoCV Ab (+) and BoCV Ab negative (-) dams, only white blood cell (WBC) values were found statistically significant (P &lt; 0.05). When the haematological parameters were compared between BoCV Ab (+)/Ag (-) and BoCV Ab (-)/Ag (-) calves, WBC (P &lt; 0.05), lymphocyte (P &lt; 0.01) and granulocyte (P &lt; 0.01) values were found statistically important. When the haematological parameters were compared between BoCV Ab (+)/Ag (+) and BoCV Ab (-)/Ag (-) calves, both lymphocyte and granulocyte values were statistically important (P &lt; 0.01).Discussion: BoCV infection has found worldwide among cattle of all ages. The disease results in major economic losses in dairy herds that result from treatment costs and calf deaths. One hundred seventy two out of 183 mothers whose blood sampling was done were detected as seropositive. Many researchers found similar results in dairy cattle. It was detected the highest seropositivity in cattle more than six years old. One hundred seventy two blood samples out of 184 calves were detected seropositive. Also, the highest seropositivity was detected among of &gt; 5 and &lt; 6 months of age. BoCV Ag (+) presence was detected in only one faecal sample of one calf out of 184. Researchers were found same or higher BoCV Ag(+) rates in faeces of diarrheic calves. In the study lymphocyte counts of seropositive cows and in Ab(+)/Ag(+) calves determined decrease. However, the counts in seropositive calves were increased. Leukocytes levels were also high in seropositive calves. Haematocrit values were decreased in seropositive cows, calves and in Ab(+)/Ag(-) calves. BoCV infections were detected at low level in diarrheic calves. But, BoCV seropositive mature and diarrheic calves were found at high levels. Haematological application methods could be used to be supportive with the serological and virological methods. All farm managements should be maintained with strict hygiene practices. Milking bottle, calf pens or hutches need to be sanitized. The calves must be prevent contamination from faeces and urine of other calves. The protective vaccination must be applied all animals.

RT-PCR specific for Cryspovirus is a highly sensitive method for detecting Cryptosporidium parvum oocysts

Cryptosporidium parvum represents a considerable health risk to humans and animals because the parasite has a low infectious dose and is usually present at low numbers in environmental samples, which makes detection problematic. The purpose of this study was to evaluate Cryspovirus as a target for sensitive detection of C. parvum in clinical samples. Semi-quantitative RT-PCR (sqRT-PCR) and quantitative RT-PCR (qRT-PCR) directed to Cryspovirus sequences could detect less than 5 Cryptosporidium oocysts in RNA extracted from C. parvum-containing calf feces. Of interest was that a similar level of sensitivity was observed using RNA present in DNA extracts of the same C. parvum fecal samples. There was a strong correlation between both the sqRT-PCR and qRT-PCR product and number of C. parvum oocysts. Analysis of DNA extracted from a similar number of oocysts using PCR targeting the Cryptosporidium SSU rDNA gene sequence found that nested PCR was necessary to obtain a detectable PCR signal. The availability of DNA allowed for Cryptosporidium genotyping based on SSU rDNA sequencing as well as C. parvum subtyping through GP60 sequencing. By using DNA that contains viral RNA, the assay avoids two separate extractions — one for RNA and one for DNA. This two-step assay, first to detect Cryptosporidium by Cryspovirus-specific RT-PCR followed by nested SSU rDNA PCR for Cryptosporidium genotyping may represent an important tool for identifying the parasite in clinical samples.

Contamination of Australian newborn calf carcasses at slaughter with Clostridium difficile

In North America and Europe, reports of a genetic overlap between toxigenic strains of Clostridium difficile isolated from humans, livestock and retail meat suggest that food-borne transmission may be occurring. We investigated the prevalence, concentration and genetic diversity of C. difficile on the carcasses (n = 300) and in the faeces (n = 30) of neonatal veal calves at three abattoirs in Australia in 2013. Selective culture (both direct and enrichment) was performed, and all isolates were characterized by PCR for the toxin genes tcdA, tcdB and cdtA/B and by PCR ribotyping. Prevalence of C. difficile was 25.3% (76/300) on carcasses and 60.0% (18/30) in faeces. Multiple PCR ribotypes (RT) were detected, with four binary toxin-positive RTs accounting for 70.3% (71/101) of isolates; 127 (A+, B+, CDT+, 32.7%), 288 (A–, B–, CDT+, 28.7%), 033 (A–, B–, CDT+, 6.9%) and 126 (A+, B+, CDT+, 2.0%). Viable counts of a subset of samples revealed detectable numbers of C. difficile in 66.7% (10/15) of faecal samples (range 2.0 × 103 to 2.3 × 106 CFU/mL, median count 2.5 × 104 CFU/mL) and in 16.7% (25/150) of carcase samples (range 3 to 33 CFU/cm2, median count 7 CFU/cm2). These data further confirm that Australian neonatal veal calf carcasses are contaminated with potentially significant strains of C. difficile at slaughter.

Sesquiterpene lactone containing extracts from two cultivars of forage chicory (Cichorium intybus) show distinctive chemical profiles and in vitro activity against Ostertagia ostertagi

The study investigated direct anthelmintic effects of sesquiterpene lactones (SL)-containing extracts from forage chicory against free-living and parasitic stages of Ostertagia ostertagi. Freeze-dried leaves from chicory cultivars ‘Spadona’ and ‘Puna II’ were extracted using methanol/water. Total SL were further fractionated by solid-phase extraction and resulting extracts were characterised by high-performance liquid chromatography (HPLC). O. ostertagi eggs from faeces of mono-infected calves were hatched and L1 were used in a larval feeding inhibition assay (LFIA), while cultured L3 were used in a larval exsheathment inhibition assay (LEIA). Adult worms were immediately recovered after slaughter and used for motility inhibition assays (AMIA). Electron microscopy (EM) was performed on adult O. ostertagi exposed to 1000 μg extract mL−1 of both chicory cultivars. In all assays, decreasing concentrations of SL-containing extracts in PBS (1% DMSO) were tested in replicates with 1% DMSO in PBS as negative controls. HPLC demonstrated similar concentrations of most SL in both extracts. However, Spadona-extract contained significantly higher concentrations of 11, 13-dihydro-8-deoxylactucin (P = 0.01), while Puna II-extract had increased levels of 11, 13-dihydrolactucin (P < 0.0001). In the LFIA, both extracts reduced larval feeding at increasing concentrations, but Spadona-extract showed higher potency confirmed by significantly lower EC50 (P < 0.0001). In the LEIA, neither of the two extracts interfered with the exsheathment of L3 (P > 0.05). In the AMIA, both SL-containing extracts induced a dose-dependent effect but Spadona-extract showed greater activity and exerted faster worm paralysis than Puna II-extract with significantly lower EC50 (P < 0.0001). No cuticular damage was observed by EM in worms exposed to any of the extracts. We have demonstrated that SL-containing extracts from forage chicory can inhibit feeding of free-living larvae and exert direct effects against parasitic stages of O. ostertagi. Our results may contribute to the identification of natural anti-parasitic compounds and to interpret the in vivo anthelmintic effects of forage chicory.