Calf Diarrhea Research Articles

Development and application of one-step multiplex Real-Time PCR for detection of three main pathogens associated with bovine neonatal diarrhea.

Neonatal calf diarrhea (NCD) is one of the most common diseases in calves, causing huge economic and productivity losses to the bovine industry worldwide. The main pathogens include bovine rotavirus (BRV), bovine coronavirus (BCoV), and Enterotoxigenic Escherichia coli (ETEC) K99. Since multiple infectious agents can be involved in calf diarrhea, detecting each causative agent by traditional methods is laborious and expensive. In this study, we developed a one-step multiplex Real-Time PCR assay to simultaneously detect BRV, BCoV, and E. coli K99+. The assay performance on field samples was evaluated on 1100 rectal swabs of diseased cattle with diarrhea symptoms and compared with the conventional gel-based RT-PCR assay detect BRV, BCoV, and E. coli K99+. The established assay could specifically detect the target pathogens without cross-reactivity with other pathogens. A single real-time PCR can detect ~1 copy/µL for each pathogen, and multiplex real-time PCR has a detection limit of 10 copies/µL. Reproducibility as measured by standard deviation and coefficient of variation were desirable. The triple real-time PCR method established in this study was compared with gel-based PT-PCR. Both methods are reasonably consistent, while the real-time PCR assay was more sensitive and could rapidly distinguish these three pathogens in one tube. Analysis of surveillance data showed that BRV and BCoV are major enteric viral pathogens accounting for calves' diarrhea in China. The established assay has excellent specificity and sensitivity and was suitable for clinical application. The robustness and high-throughput performance of the developed assay make it a powerful tool in diagnostic applications and calf diarrhea research. ​.

Evaluation of serum concentrations of total protein and gamma-globulin as an indicator of serum immunoglobulin G concentration in dairy calves

The aim of this study was to evaluate precision of estimating serum immunoglobulin G (IgG) concentration from total protein (TP) or gamma-globulin (γGLB) concentration as an alternative approach, and to compare morbidity of pre-weaned dairy calves differing in serum γGLB concentration. In trial 1, blood was sampled from 129 Holstein calves in the first week after birth, and serum concentrations of TP, γGLB, and IgG were measured. Spearman's correlation coefficient (rs) between serum IgG and TP concentrations was 0.89, and rs between serum IgG and γGLB concentrations was 0.96. Absolute residual (observed – predicted) serum IgG concentrations were smaller when estimated by serum γGLB concentration than by serum TP concentration, and differences in the absolute residuals were smaller for calves fed colostrum replacer (1.68 vs. 4.29 g/L) than those fed whole colostrum (2.41 vs. 3.48 g/L). In trial 2, blood was sampled from 740 Holstein heifer calves during the first week of age, and serum γGLB concentration was measured. The calves were divided into 4 categories based on their serum γGLB concentration; ≥ 1.0 g/dL (Excellent), 0.7 ≤ γGLB <1.0 g/dL (Good), 0.4 ≤ γGLB <0.7 g/dL (Fair) and <0.4 g/dL (Poor). Morbidity for diarrhea and respiratory disease in pre-weaned dairy calves was determined for the first 28 and 56 d of age, and compared among the 4 categories based on serum γGLB concentration. Calves with serum γGLB concentration higher than 0.7 g/dL (Good and Excellent) had less diarrhea during the first 28 d of age than those with lower serum γGLB concentration (Fair and Poor). Calves with serum γGLB concentration higher than 1.0 g/dL (Excellent) had less respiratory diseases for the first 56 d of age than those with lower serum γGLB concentration (Good, Fair, and Poor). These results suggest that serum IgG concentration can be estimated more precisely from concentration of γGLB than TP particularly for calves fed colostrum replacer, and that γGLB concentrations in the first week of age are associated with morbidity of calves. Transfer of passive immunity in dairy calves can be assessed effectively by serum γGLB concentration.

Comprehensive genetic analysis of the first near-complete genome of bovine coronavirus and partial genome of bovine rotavirus in Türkiye through metagenomics.

Obtaining the complete or near-complete genome sequence of pathogens is becoming increasingly crucial for epidemiology, virology, clinical science and practice. This study aimed to detect viruses and conduct genetic characterization of genomes using metagenomics in order to identify the viral agents responsible for a calf's diarrhoea. The findings showed that bovine coronavirus (BCoV) and bovine rotavirus (BRV) are the primary viral agents responsible for the calf's diarrhoea. The current study successfully obtained the first-ever near-complete genome sequence of a bovine coronavirus (BCoV) from Türkiye. The G+C content was 36.31% and the genetic analysis revealed that the Turkish BCoV strain is closely related to respiratory BCoV strains from France and Ireland, with high nucleotide sequence and amino acid identity and similarity. In the present study, analysis of the S protein of the Turkish BCoV strain revealed the presence of 13 amino acid insertions, one of which was found to be shared with the French respiratory BCoV. The study also identified a BRV strain through metagenomic analysis and detected multiple mutations within the structural and non-structural proteins of the BRV strain, suggesting that the BRV Kirikkale strain may serve as an ancestor for reassortants with interspecies transmission, especially involving rotaviruses that infect rabbits and giraffes.

Management of bacterial neonatal diarrhea in calves

Diseases such as neonatal diarrhea are common in poorly managed herds and can cause major losses in any cattle breeding system in the world, especially in intensive systems, accounting for 2% of herd losses. As soon as the calf is born, it is exposed to numerous factors, such as agents, the host and the environment, which together can favor the onset of this disease because, due to the reproductive characteristics of the female bovine, there is no transfer of transplacental immunity to the newborn.

Identification of major protozoal enteropathogens causing calf diarrhea in dairy farms in and around Holeta Town, Oromia Special Zone, Ethiopia

Calf diarrhea can be caused by a variety of pathogens, including bacteria, viruses, protozoa, and intestinal parasites. Giardia, Eimeria, and Cryptosporidium are the most significant protozoan parasites and are all individually and ollectively infectious. A case-series study was carried out in and around Holeta dairy farms, Oromia Special Zone, Ethiopia, from November 2017 to April 2018 to identify the main protozoan enteropathogens from diarrheic calves. Samples were purposely collected from three dairy farms: Serkalem Dairy Farm (SDF), Holeta Agricultural Research Centre (HARC), and Ada Berga Agricultural Research Centre (ABARC). A total of 93 fecal samples were taken from diarrheic calves of up to 4 months of age. Protozoan enteropathogens were identified using flotation and modified Ziehl Neelsen staining methods. Cryptosporidium, Eimeria, and Giardia were detected in 44 (47.3%), 50 (53.8%), and 34 (36.6%) of the diarrheic fecal samples examined, respectively. The findings indicated that there were 17 (18.27%) cases of Cryptosporidium, Eimeria, and Giardia as mixed infections, 16 (17.2%) cases of Cryptosporidium and Eimeria, 5 (5.4%) cases of Cryptosporidium and Giardia, and 4 (4.3%) cases of Eimeria and Giardia, compared to 13 (13.97%), 6 (6.45%), and 5 (5.4%) cases of Cryptosporidium, Eimeria and Giardia as single infections, respectively. The selected farms had significant prevalence levels of three common protozoa: Eimeria, Giardia, and Cryptosporidium. Further molecular research is required to identify the species and genotype levels of protozoal enteropathogens and related risk factors.

Association of passive immunity and genetic composition, health, and performance of tropical dairy calves

This retrospective cohort study aimed to identify risk factors for the failure of transfer of passive immunity (FPI) in preweaned dairy calves, explore its associations with morbidity, mortality, genetics, and determine a standardized cut-off point for FPI. Analyzing data from 6011 calves, factors such as sire predicted transmitted ability for milk (PTA), birth season, retained placenta (RP), total serum protein concentration (TSP), morbidity (neonatal calf diarrhea - NCD, bovine respiratory disease - BRD, tick-borne disease - TBD), mortality, and average daily weight gain (ADG) were considered. The calves were categorized into predominantly Gyr (PG) and predominantly Holstein (PH) genetic composition groups. Multivariate mixed logistic regression revealed optimal TSP cut-off points for predicting morbidity (7.6 g/dL) and mortality (6.9 g/dL). PH calves exhibited 1.35 times higher odds of FPI and 1.48 times greater odds of disease. Calves from multiparous cows and those born to dams with RP had increased FPI odds. Disease prevalence was 53%, with 41% NCD, 18% BRD, and 10% TBD. Season, parity, PTA, and birth weight were associated with disease odds, though FPI was not a reliable predictor. The mortality percentage was 6%, with PH calves and those with a positive PTA having higher odds. The ADG was 0.64 kg, and FPI-affected calves gained less weight. The study compared and identified various risk factors that potentially impact calf immunity. However, the use of a standardized cut-off point for FPI assessment was not effective in predicting morbidity and mortality at this specific farm.

Bovine neonatal enteric immunity: a key consideration in the prevention of calf diarrhoea

The mucosa is the largest component of the immune system, providing a first line of defence against most pathogens encountered. Although the mucosa is located in all areas where there are mucosal surfaces (such as the respiratory and urogenital tracts), the clinical importance of diarrhoea in calf veterinary practice means the gastrointestinal mucosa is of particular interest. Taking steps to promote good health of the gastrointestinal mucosa can enhance gastrointestinal mucosal immunity and aid prevention of calf diarrhoea, the leading cause of mortality and morbidity in calves under 1 month old.

140 High-resolution characterization of the gut microbiota of neonatal dairy calves using full-length 16S rRNA gene amplicon sequencing

Abstract The gut microbiota has a crucial role in preventing pathogen colonization and training the immune system. An imbalanced gut microbiota tends to induce calf diarrhea, which can lead to dehydration and metabolic acidosis. Therefore, it is urgent to identify critical factors that influence the establishment of gut microbiota in the early stages of development in dairy calves. Most previous research relied on short-read-based partial 16S rRNA gene amplicon sequencing to study calf microbiota, which has limited taxonomic resolution. The aim of our study is to obtain a species-level characterization of calf gut microbiota using long-read based full-length 16S rRNA gene amplicon sequencing and investigate the factors that affect the gut microbiota composition in the early stages of development in dairy calves. We collected fecal samples from the recto-anal junction of 25 Holstein-Angus crossed calves at d 4 and d 14 after birth. The calves were raised in individual outdoor hutches from birth and fed milk replacer twice daily. The gut microbiota was analyzed using full-length 16S rRNA gene amplicon sequencing on an Oxford Nanopore sequencer. The results revealed that the gut microbial diversity measured by Chao 1 (P &amp;lt; 0.001) and Shannon index (P &amp;lt; 0.001) significantly increased from d 4 to d 14. We identified 15 and 36 core bacterial species (prevalence ≥ 80%) in d 4 and d 14 gut microbiota, respectively, and both stages share 7 bacterial species: Bacteroides fragilis, Butyricicoccus pulicaecorum, Enterocloster bolteae, Escherichia coli, Lachnoclostridium sp YL32, Rumiococcus gnavus and Streptococcus equinus. The calf gut microbiota was dominated by Bacteroides fragilis (18%), Escherichia coli (13%), Ruminococcus gnavus (10%) and Butyricicoccus pulicaecorum (6%) species on d 4, and Megamonas rupellensis (18%) and Fusobacterium varium (6%) species (relative abundance &amp;gt; 5%) on d 14. Notably, we found a significant impact of hutch location on the composition of gut microbiota, particularly on d 4, but the impact was reduced on d 14. Moreover, there were 4 dystocia calves among the 25 calves, which had a significantly lower Shannon index (P = 0.046) compared with 4 paired normal calves of similar age and sex, and location balanced. In conclusion, this study provides a species-level resolution of gut microbiota for neonatal dairy calves and indicates a potential impact of dystocia on early microbiota development. Additionally, calf microbiota composition is influenced by environmental factors, likely the soil microbiota where hutches were located, with a more substantial effect on the microbiota at an earlier age.

The first complete genome sequence and genetic evolution analysis of bovine norovirus in Xinjiang, China

Abstract Introduction Viruses are among the main pathogens causing diarrhoea in calves. The current study found that bovine norovirus (BNoV) is one of the principal viruses causing diarrhoea in calves in Xinjiang, China. Material and Methods A total of 974 calf faecal samples from six regions in Xinjiang were tested for BNoV using reverse-transcriptase PCR. The genomic characteristics of BNoV and the genetic evolution of the VP1 gene, protein three-dimensional structure characteristics and amino acid variation were analysed using bioinformatics methods. Results Epidemiological survey results showed that the infection rate of BNoV was 19.82%, and all samples tested positive in five regions. The results of the genetic evolution analysis showed that BNoV strains from Tacheng of northern Xinjiang and Kashgar of southern Xinjiang both belonged to the GIII.2 genotype of BNoV but were not on the same cluster of evolutionary branches. Additionally, the amino acid variation of the VP1 protein was not observed to significantly affect its spatial structure. Conclusion This study is the first to report the genetic characteristics of the BNoV complete genome sequence in Xinjiang and provides a scientific basis for BNoV vaccine development and pathogenesis research.

Enterobacteria Isolated From Diarrheic Calves and their Phenotypic Resistance Pattern

Abstract Diarrhea is a prevalent and economically impactful ailment in cattle herds, resulting in substantial losses attributed to mortality, treatment expenses, and impaired calf growth. The aim of this study was to establish data about the occurrence of the main bacterial agents involved in diarrhea in dairy calves in the state of Maranhão, as well as to determine the most effective and/or resistant antibiotics, establishing a foundation for treatment and prevention protocols. The samples were collected with rectal swabs and initially cultivated on sheep blood agar (5%) and MacConkey agar and later on Salmonella-Shigella agar and eosin-methylene blue agar. Gram staining, biochemical tests, and antibiograms were performed on the obtained colonies. Ten municipalities were studied, with 230 calves evaluated from ten dairy farms. A total of 21 animals exhibited clinical signs of diarrhea, representing a frequency of 9.13%. The frequency of positive farms was 35%, and in 50% of municipalities. Bacteria isolated belonged to the Enterobacteriaceae family, with 71.4% Escherichia coli, 14.3% Enterobacter sp., 9.5% Proteus sp. and 4.8% Klebsiella sp. On the antibiogram, the bacteria that showed the highest resistance levels were those of the Proteus genus, followed by Enterobacter sp. and E. coli. The bacteria found are relevant for both animal and human health due to their zoonotic potential and serve as a public health alert since the isolates in this study showed in vitro resistance to several antibiotics, which predisposes them to the emergence of multidrug-resistant bacteria.

The first commercially approved efficacious cryptosporidium vaccine protecting New-Born calves from severe diarrhea

Neonatal calf diarrhea (NCD) is an important cause of morbidity and mortality in calves. Four major pathogens, including Enterotoxigenic Escherichia coli (ETEC), bovine rotavirus (BRV), bovine coronavirus (BCV), and Cryptosporidium spp., are associated with NCD, with Cryptosporidium parvum (C. parvum) recognized as a major contributor. Despite various treatment options available, there is no effective vaccine for bovine cryptosporidiosis yet. In this study, recombinantly expressed C. parvum gp40 was evaluated as a candidate vaccine in pregnant animals. The vaccine was administered to pregnant cows during the last trimester of their pregnancy resulting in highly statistically significantly increased anti-gp40 antibody titers. The resulting anti-gp40 antibodies were transmitted to the calves via colostrum. Both suckling (n = 29; test:14, control:15) and non-suckling (n = 16; 8 per group) calves were challenged with live C. parvum oocysts. The calves that received the colostrum from gp40 vaccinated cows showed a highly statistically significant improvement in health (p < 0.0001), reduced incidence (p < 0.0001), and duration of severe diarrhea (p < 0.001) and increased weight gain in suckling calves (p < 0.0001). This study demonstrates that immune bovine colostrum, induced by immunization of late-gestation cows with C. parvum gp40, provided substantial protection to calves against cryptosporidiosis. These findings suggest that the gp40 vaccine has potential for preventing outbreaks of neonatal diarrhea associated with cryptosporidiosis in calves, thereby improving animal welfare and performance.

Neonatal Calf Diarrhea and Gastrointestinal Microbiota: Etiologic Agents and Microbiota Manipulation for Treatment and Prevention of Diarrhea.

Neonatal calf diarrhea is the leading cause of neonatal morbidity and mortality globally. The changes associated with the gastrointestinal microbiota in neonatal calves experiencing diarrhea and its etiology are not fully understood or completely defined in the literature. Several studies have demonstrated that the fecal microbiota of calves that experience diarrhea substantially deviates from that of healthy age-matched calves. However, one key question remains: whether the changes observed in the bacterial communities (also known as dysbiosis) are a predisposing factor for, or the consequence of, gastrointestinal inflammation caused by the pathogens associated with calf diarrhea. The first objective of this literature review is to present the current information regarding the changes in the fecal microbiota of diarrheic calves and the impact of the pathogens associated with diarrhea on fecal microbiota. Modulation of the gastrointestinal microbiota using pre- and probiotics, colostrum feeding, and fecal microbiota transplantation (FMT) has been used to treat and prevent gastrointestinal diseases in humans and dogs. Although information regarding the use of probiotics for the prevention of diarrhea is available in cattle, little information is available regarding the use of these strategies for treating calf diarrhea and the use of prebiotics or FMT to prevent diarrhea. The second objective of this literature review is to summarize the current knowledge regarding the impact of prebiotics, probiotics, synbiotics, colostrum feeding, and FMT for the treatment and prevention of calf diarrhea.

Exploring genetic polymorphisms and transcript levels of antioxidant and metabolic markers for prediction and monitoring diarrhea in Holstein dairy calves

ABSTRACT The objective of this study was to investigate the genetic polymorphisms and transcript levels of antioxidant and metabolic markers for diarrhea monitoring in Holstein dairy calves. For DNA and RNA extraction, blood samples were taken from 100 female calves (50 with diarrhea and 50 appearing healthy). DNA sequencing and real time PCR for and metabolic genes were performed to identify genetic variants linked to diarrhea susceptibility. Results revealed that there were nucleotide sequence variations in the investigated genes between healthy and diarrheal calves. Keap1, HMOX1, CMPK2, ASPG, FPGT, TNNI3 K, and LPCAT1 were considerably more highly expressed in diarrheic calves than resistant ones. The Nrf2, PRDX2, and PRDX6 genes, however, produced a different pattern. This study provides a practical control strategy to lessen newborn diarrhea in Holstein dairy calves and emphasizes the significance of nucleotide variations along with antioxidant and metabolic gene expression patterns as proxy markers of diarrhea susceptibility/resistance.

Differential Microbial Composition and Interkingdom Interactions in the Intestinal Microbiota of Holstein and German Simmental × Holstein Cross F1 Calves: A Comprehensive Analysis of Bacterial and Fungal Diversity.

Calf intestines are colonized by rich and complex microbial communities, playing a crucial role in animal physiology, metabolism, nutrition, and immune function. In this study, we provide insight into the composition of fecal microbial bacteria and fungi, respectively, as well as the cross-kingdom interactions. We investigated the intestinal microbiota of different breeds of calves by characterizing the bacterial and fungal communities in the rectal feces of Holstein calves and German Simmental × Holstein cross F1 generation (GXH) using 16S rRNA and ITS amplicon sequencing techniques. PICRUSt2 (version 2.2.0) were used to determine microbial diversity and function and explore the reasons why Holstein calves are more susceptible to diarrhea. The results revealed no significant difference in the diversity of fecal microbiota among the groups (p > 0.05). We identified Firmicutes, Bacteroidetes, and Proteobacteria as the dominant bacterial phyla in the fecal bacterial communities of the two breeds of calves. Ascomycota and Basidiomycota play important roles in the fungal community but differ in relative abundance. Bacteroides was the dominant genus at the group level for calf fecal microbiota in both breeds. The relative abundance of Prevotella, Escherichia-Shigella, Peptostreptococcus, and Butyricicoccus was higher in Holstein calves, and the relative abundance of Faecalibacterium, Megamonas, Butyricicoccus, and Alloprevotella was lower than GXH group. Aspergillus and Cladosporium were the dominating genera of fecal fungi in both groups of calves. LEfSe analysis revealed 33 different bacteria and 23 different fungi between the two groups, with more differential strains found in GXH. In addition, the feces fungi-bacteria interkingdom interactions varied among breeds. Thus, the composition and structure of bacterial and fungal communities in calf feces varied by breed, indicating a potential association between breed and microbial communities. We also found differences in the network between bacterial-fungal kingdoms. We explain the reasons for Holstein calves being more prone to diarrhea. This indicated that breed makes differences in calf diarrhea rates by influencing gut microbial composition and interactions.

Sodium acetate and sodium butyrate attenuate diarrhea in yak calves by regulating gut microbiota and metabolites

Diarrhea is a severe issue in calves that causes fertility problems and economic issues worldwide. Sodium acetate/sodium butyrate (SA/SB) alleviates diarrhea in mice; however, little information is available about the preventive effect of SA/SB on diarrheic yak calves living on the Tibet plateau. Yak calves (n = 19) of age ≥4 months and weight 37 ± 2 Kg were randomly divided into control (C, n = 10) and supplement groups (S, n = 9). Yaks belonging to the supplement group were given sodium butyrate (10 g/kg) and sodium acetate (5 g/kg) for 28 days, along with normal feed, seasonal grasses, pasture, and water. The blood and fecal samples from yak calves were collected for assessment of antioxidant capacity, inflammatory cytokines, microbiome, and short-chain fatty acids (SCFAs) concentration analysis. Results of this study revealed that a lower diarrhea rate, higher weight, and net weight gain were recorded in yaks belonging to group S supplemented with SA/SB. Similarly, increased antioxidant capacity with higher levels of T-AOC, SOD, and GSH-px and decreased inflammatory reactions by decreasing both TNF-α and IL-1β concentrations were recorded in yaks of group S. The concentration of SCFAs was significantly higher (p < 0.05) in yaks from group S than group C. Microbiome analysis revealed that 8 phyla and 54 genera were significantly different (p < 0.05) in both yak groups, with increased probiotics (Akkermansia, Oscillospira), SCFAs producing genera (Oscillospira, ASF356, Anaerosporobacter and Phascolarctobacterium), and decreased inflammatory related genus (Flavonifractor, Fournierella) and harmful bacteria (Oscillibacter, Achromobacter) in group S. In conclusion, the results demonstrated that SA and SB could decrease diarrhea rates in yak calves on the plateau via increasing antioxidant ability and SCFAs, while decreasing inflammatory responses in yaks by moderating gut microbiota. The current results provide new insights for the prevention and treatment of diarrhea in yaks.

Impact of zinc supplementation on phenotypic antimicrobial resistance of fecal commensal bacteria from pre-weaned dairy calves.

The objective of this study was to evaluate the impact of dietary zinc supplementation in pre-weaned dairy calves on thephenotypic antimicrobial resistance (AMR) of fecal commensal bacteria. A repository of fecal specimens from a random sample of calves block-randomized into placebo (n = 39) and zinc sulfate (n = 28) groups collected over a zinc supplementation clinical trial at the onset of calf diarrhea, calf diarrheal cure, and the last day of 14 cumulative days of zinc or placebo treatment were analyzed. Antimicrobial susceptibility testing was conducted for Enterococcus spp. (n = 167) and E. coli (n = 44), with one representative isolate of each commensal bacteria tested per sample. Parametric survival interval regression models were constructed to evaluate the association between zinc treatment and phenotypic AMR, with exponentiated accelerated failuretime (AFT) coefficients adapted for MIC instead of time representing the degree of change in AMR (MIC Ratio, MR). Findings from our study indicated that zinc supplementation did not significantly alter the MIC in Enterococcus spp. for 13 drugs: gentamicin, vancomycin, ciprofloxacin, erythromycin, penicillin, nitrofurantoin, linezolid, quinupristin/dalfopristin, tylosin tartrate, streptomycin, daptomycin, chloramphenicol, and tigecycline (MR = 0.96-2.94, p > 0.05). In E. coli, zinc supplementation was not associated with resistance to azithromycin (MR = 0.80, p > 0.05) and ceftriaxone (MR = 0.95, p > 0.05). However, a significant reduction in E. coli MIC values was observed for ciprofloxacin (MR = 0.17, 95% CI 0.03-0.97) and nalidixic acid (MR = 0.28, 95% CI 0.15-0.53) for zinc-treated compared to placebo-treated calves. Alongside predictions of MIC values generated from these 17 AFT models, findings from this study corroborate the influence of age and antimicrobial exposure on phenotypic AMR.

Clinical Insights of Lumpy Skin Disease in Cauvery Delta Region of Tamil Nadu

Lumpy Skin Disease (LSD) is a contagious and life-threatening affliction affecting both cattle and calves, marked by the emergence of nodules on the skin and various body areas. Notably, in calves, the impact extends beyond generalized lesions, leading to a frequent occurrence of respiratory distress. Secondary bacterial infection also often aggravates the condition. A total of 36 cases presented to Veterinary Clinical Complex, Veterinary College and Research Institute, Orathanadu during the period of April to June 2023 with the history of eruptive lesions all over the body showing signs of Lumpy skin disease with various clinical manifestations like nodular lesions all over the body, fever, nasal discharge, lymphadenopathy, inspiratory dyspnoea, along with diffused edematous swelling of all limbs and diarrhoea in calves. In Indigenous adult cattle, diffused edematous swelling of limbs either unilateral or bilateral along with brisket and jowl edema were observed. Mortality rate in calves less than 3 months is 75 %. In adult animal the mortality rate was high in Indigenous breeds compared to cross bred cattle. Animals were treated to combat secondary bacterial infections with antibiotics along with supportive like Syr. Lavitone H 3ml per.os daily as prophylactic therapy to combat fungal pneumonia. The cases with severe clinical manifestation were even treated with steroid Inj. Prednisolone @1mg/kg b.wt. Animal treated for 15 days, gradual recovery was noticed. Vector control need to be implemented to prevent the disease associated with economic losses.

Effect of Dietary Supplementation of Chestnut and Quebracho Tannin Supplementation on Neonatal Diarrhoea in Preweaning Calves.

Neonatal calf diarrhoea (NCD) poses a significant health challenge in cattle herds, resulting in considerable economic losses and antimicrobial use. In response to the escalating threat of antimicrobial resistance, viable alternatives are imperative, aligning with European policies. This study evaluated the in-milk supplementation of the chestnut and quebracho tannin extract in preweaning calves on performance, diarrhoea occurrence, Cryptosporidium spp. shedding, protein digestibility, and intestinal health. Twenty newborn calves were divided, after colostrum administration, into two experimental groups for 30 days as follows: the control (CTRL) was fed with whole milk and solid feed, and tannins (TAN) were fed whole milk supplemented with 6/g day of tannin extract and solid feed. Faecal samples were collected on days 0, 3, 7, 14, and 30 for the evaluation of Cryptosporidium oocyst shedding and protein digestibility. Faecal consistency was evaluated during the sampling using the faecal score scale (0-3 scale, considering diarrhoea > 1). The results showed a significant reduction in diarrhoea frequency in the TAN compared to the CTRL group (p < 0.05) over 30 days of the trial. The prevalence of Cryptosporidium spp. was generally low (12%), considering all analysed samples. Protein digestibility revealed comparable values for the TAN and CTRL groups, suggesting that tannins did not negatively affect milk protein availability. In conclusion, the in-milk supplementation of 6/g day of the chestnut and quebracho tannin extract could be considered a valuable functional feed additive to decrease NCD occurrence, thus supporting animal health and decreasing antibiotic use in livestock.

Effects of a commercial feed additive used for prophylactic purposes on health and blood parameters in neonatal calves.

Neonatal calf diarrhea is commonly observed worldwide and leads to significant economic losses. The objective of this study was to investigate the effect of a feed additive supplemented with milk on the incidence of diarrhea days with scours, fecal score, average daily gain, fecal bacterial concentration, and blood parameters in neonatal calves. Forty Holstein-Friesian crossbreed (HF x Montbeliarde) calves were divided into control (C) and experimental (E) groups, each containing 20 calves. The study was conducted over a period of 10 days. The control group was fed whole milk twice daily. The experimental group was fed the same whole milk supplemented with feed additive once in the morning at 100 g/calf/day. At the end of the study, it was determined that the feed additive in the milk affected fecal score, blood pH, pCO2, cHCO3 -, K+, cHgb, and plasma glucose levels (p<0.05), but did not affect the incidence of diarrhea, average daily gain, days with scours, and other blood parameters (p>0.05). Rotavirus was the main pathogen found in the fecal samples of calves with diarrhea. Higher concentrations of Escherichia coli, Salmonella spp., and Clostridium perfringens were detected in the feces of experimental calves (p<0.001). The findings indicate that this feed additive improves fecal score, some blood parameters, and fecal concentration of pathogenic bacteria in feces. Additionally, it shows a favorable effect on both diarrhea incidence and the days with scours, suggesting its potential efficacy in preventing diarrhea in neonatal calves. Preventing diarrhea during this crucial stage of life can contribute to the health and productivity of animals and play a role in preventing antibiotic resistance by reducing reliance on antibiotics.

Characterization and Spike Gene Analysis of a Candidate Attenuated Live Bovine Coronavirus Vaccine

The bovine coronavirus (BCoV) KBR-1 strain, obtained from calf diarrhea samples collected in 2017, belongs to group GIIa. To attenuate this strain, it was subcultured continuously (up to 79 times) in HRT-18 cells, followed by 80–120 passages in MDBK cells. The KBR-1-p120 strain harvested from MDBK cells at passage 120 harbored 13 amino acid mutations in the spike gene. Additionally, the KBR-1-p120 strain showed a high viral titer and cytopathogenic effects in MDBK cells. Seven-day-old calves (negative for BCoV antigen and antibodies) that did not consume colostrum were orally inoculated with the attenuated candidate strain (KBR-1-p120), or with KBR-1 passaged 10 times (KBR-1-p10) in HRT-18 cells. Calves inoculated with KBR-1-p10 had a low diarrhea score, and BCoV RNA was detected at 3–7 days post-inoculation (DPI). The virus was also present in the duodenum, jejunum, and ileum at autopsy; however, calves inoculated with KBR-1-p120 had low levels of BCoV RNA in feces at 4–6 DPI, and no diarrhea. In addition, an extremely small amount of BCoV RNA was present in the jejunum and ileum at autopsy. The small intestines of calves inoculated with KBR-1-p120 were emulsified and used to infect calves two more times, but pathogenicity was not recovered. Therefore, the KBR-1-p120 strain has potential as a live vaccine candidate.