Unveiling genetic signatures associated with resilience to neonatal diarrhea in lambs through two GWAS approaches.

Neonatal diarrhea presents a significant global challenge due to its multifactorial etiology, resulting in high morbidity and mortality rates, and substantial economic losses. While molecular-level studies on genetic resilience/susceptibility to neonatal diarrhea in farm animals are scarce, prior observations indicate promising research directions. Thus, the present study utilizes two genome-wide association approaches, pKWmEB and MLM, to explore potential links between genetic variations in innate immunity and neonatal diarrhea in Karacabey Merino lambs. Analyzing 707 lambs, including 180 cases and 527 controls, revealed an overall prevalence rate of 25.5%. The pKWmEB analysis identified 13 significant SNPs exceeding the threshold of ≥ LOD 3. Moreover, MLM detected one SNP (s61781.1) in the SLC22A8 gene (p-value, 1.85eE-7), which was co-detected by both methods. A McNemar's test was conducted as the final assessment to identify whether there are any major effective markers among the detected SNPs. Results indicate that four markers-oar3_OAR1_122352257, OAR17_77709936.1, oar3_OAR18_17278638, and s61781.1-have a substantial impact on neonatal diarrhea prevalence (odds ratio: 2.03 to 3.10; statistical power: 0.88 to 0.99). Therefore, we propose the annotated genes harboring three of the associated markers, TIAM1, YDJC, and SLC22A8, as candidate major genes for selective breeding against neonatal diarrhea.

Diet management in congenital diarrheas and enteropathies - general concepts and disease-specific approach, a narrative review.

Congenital diarrheas and enteropathies (CODE) are a group of rare, heterogenous, monogenic disorders that lead to chronic diarrhea in infancy. Definitive treatment is rarely available, and supportive treatment is the mainstay. Nutritional management in the form of either specialized formulas, restrictive diet, or parenteral nutrition support in CODE with poor enteral tolerance is the cornerstone of CODE treatment and long-term growth. The evidence to support the use of specific diet regimens and nutritional approaches in most CODE disorders is limited due to the rarity of these diseases and the scant published clinical experience. The goal of this review was to create a comprehensive guide for nutritional management in CODE, based on the currently available literature, disease mechanism, and the PediCODE group experience. Enteral diet management in CODE can be divided into 3 distinct conceptual frameworks: nutrient elimination, nutrient supplementation, and generalized nutrient restriction. Response to nutrient elimination or supplementation can lead to resolution or significant improvement in the chronic diarrhea of CODE and resumption of normal growth. This pattern can be seen in CODE due to carbohydrate malabsorption, defects in fat absorption, and occasionally in electrolyte transport defects. In contrast, general diet restriction is mainly supportive. However, occasionally it allows parenteral nutrition weaning or reduction over time, mainly in enteroendocrine defects and rarely in epithelial trafficking and polarity defects. Further research is required to better elucidate the role of diet in the treatment of CODE and the appropriate diet management for each disease.

The vaccination changed the profile of rotavirus infection with the increase of non-rotavirus A species diagnosis in one-week-old diarrheic piglets.

Porcine rotavirus (RV) is a major viral agent associated with severe diarrhea in newborn piglets. RVA, RVB, RVC, and RVH are RV species that have already been identified in pigs. RVA is considered the most prevalent and relevant virus in pig production worldwide. This study aimed to evaluate the frequency of RV infection associated with diarrhea in suckling piglets from regular RVA-vaccinated Brazilian pig herds between 2015 and 2021. Therefore, 511 diarrheic fecal samples were collected from suckling piglets aged up to 3 weeks from 112 pig farms located in three main Brazilian pork production regions. All piglets were born to RVA-vaccinated sows. The nucleic acids of RVA, RVC, and RVH were investigated by RT-PCR assays and RVB by semi-nested RT-PCR assay. Of the diarrheic fecal samples analyzed, 221/511 (43.3%) were positive for at least one of the RV species. Regarding the distribution of RV species among the positive fecal samples that presented with only one RV species, 99 (44.8%), 63 (28.5%), and 45 (20.4%) were identified as RVB, RVC, and RVA, respectively. RVH was not identified in diarrheic piglets with a single infection. More than one RV species was identified in 14/221 (6.3%) of the diarrheic fecal samples evaluated. Co-detection of RVB + RVH (11/221; 5.0%), RVA + RVB (1/221; 0.4%), RVA + RVC (1/221; 0.4%), and RVB + RVC (1/221; 0.4%) was identified in fecal samples. The results demonstrated a significant increase in the RVC and, mainly, RVB detection rates in single infections. This study allowed us to characterize the importance of other RV species, in addition to RVA, in the etiology of neonatal diarrhea in piglets from pig herds with a regular vaccination program for RVA diarrhea control and prophylaxis.

The Effects of Milk Replacer Supplemented with Ascophyllum nodosum as a Novel Ingredient to Prevent Neonatal Diarrhea in Dairy Calves and Improve Their Health Status.

Nutrition and health during pre-weaning affect the calves' future fertility, calving age, production, and carrier length. Calves are highly susceptible to neonatal calf diarrhea (NCD), which can be fatal. NCD is due to hypovolemia and acidosis, which may involve anorexia and ataxia. The One Health principle calls for a drastic reduction in antimicrobial use. One approach is to improve animal health and reduce the use of antibiotics and functional ingredients that have beneficial effects due to bioactive compounds. Several functional ingredients and additives can be considered, and, in particular for this study, Ascophyllum nodosum was considered. The present study aimed to evaluate the role of A. nodosum as a functional ingredient implemented into the milk replacer in neonatal calves. Twelve pre-weaned Holstein Frisian calves, housed in twelve individual pens in the same environmental conditions, were divided into two groups of six animals: a control group (CTRL, n = 6) fed with a milk replacer, and a treatment group receiving milk enriched with 10 g of A. nodosum in their diet (TRT, n = 6) for 42 days. The fecal score was evaluated daily (3-0 scale) to monitor the incidence of diarrhea in the two groups. The body weight was evaluated weekly, and every two weeks feces were collected for microbiological evaluation using a selective medium for plate counting of total, lactic acid, and coliform bacteria. To verify the presence of Lactobacillus, Bifidobacterium, and Escherichia coli, real-time qPCR was used. At the beginning and at the end of the trial, blood samples were obtained for serum metabolite analysis. The growth performance did not differ in either of the two groups, but significant differences were observed in the incidence of moderate diarrhea (p-value < 0.0113), where the TRT group showed a lower incidence of cases during the 42-day period. Serum analysis highlighted higher contents of albumin, calcium, phosphorus, and total cholesterol in the TRT group compared to CTRL (p-value < 0.05). In conclusion, implementation of A. nodosum in the diet of calves can lead to better animal welfare and may reduce the use of antibiotics.

Protective efficacy of a recombinant enterotoxin antigen in a maternal immunization model and the inhibition of specific maternal antibodies to neonatal oral vaccination.

ETEC (enterotoxigenic Escherichia coli) infection is the leading cause of profuse watery diarrhea in mammals, especially among pre-weaning and post-weaning piglets in swine industry. Maternal immunization is a current rational strategy for providing protection to susceptive piglets and reducing the incidence of ETEC-associated diarrhea. Here we evaluated the protective efficiency of a recombinant antigen (MBP-SLS) fused by major enterotoxin subunits (STa-LTB-STb) via a maternal immunization model, and the impacts of maternal antibodies to neonatal oral vaccination were also investigated in the neonates. The high titers of specific IgG and sIgA in pups shown that the maternal antibodies could be transferred passively. Furthermore, the increases of IL-6 and IL-10 cytokines in breast milk and pup serum indicated that immunization on mother could effectively boost the immune system of neonates. Newborn rats from immunized mothers showed a 70% survival rate after ETEC infection. However, the mucosal immune responses of neonates were inhibited by the high level of maternal specific antibodies. Among the oral-vaccinated neonates, born from mock-immunized rats reached the highest survival after ETEC challenge. Collectively, the fusion MBP-SLS antigen could effectively induce strong immune responses in rats during pregnancy and the pups could receive passive protection through specific antibodies transferred via milk and placenta. However, the specific maternal antibodies exhibited an inhibition effect on the mucosal immune responses in offspring.

A scoping review of neonatal calf diarrhea case definitions.

Various case descriptions and scoring systems have been used to define neonatal calf diarrhea (NCD) and diverse diarrhea-related outcomes are reported, which limits direct comparison between studies. Therefore, the objective of this scoping review was to characterize the case definitions used for NCD and describe diarrhea-related outcomes to inform future efforts towards standardization. A literature search identified articles using 3 databases (Medline, CAB Direct, Agricola), along with Google and Google Scholar. This returned 16,854 unique articles, which were then screened for eligibility by two independent reviewers, resulting in 555 being selected for data extraction. Among articles, the study populations included mostly dairy-breed calves (88%; n = 486) while the remainder evaluated beef, crossbred, or dual purpose beef/dairy calves (10%; n = 53), or did not report breed (3%; n = 16). Studies used between 1 and 8 metrics to define NCD, with 933 unique metrics extracted in total. The most common metric was fecal consistency alone (30%; n = 281), or with at least 1 other metric (26%; n = 241). To define diarrhea, fecal consistency was either described qualitatively (e.g., "profuse liquid feces"), or semi-quantitatively, for example using a scoring system that frequently included 4 levels (n = 208). Some NCD case definitions included fecal color, volume, or odor (10%; n = 98), physical exam parameters (8%; n = 79), the duration of abnormal feces (7%; n = 67), the presence of abnormal contents (e.g., blood, 7%; n = 61), farm treatment records (6%; n = 54), fecal dry matter (1%; n = 12), or another metric (4%; n = 41). One or more references were cited for the NCD case definition by 49% of studies (n = 273/555), with the most common references being Larson et al. (1977) (n = 85), and McGuirk (2008) (n = 59). In the 555 included articles, 979 unique diarrhea-related outcomes were found, most commonly a binary categorization of calves having or not having diarrhea (49%; n = 483). Other articles reported statistical outcomes calculated from fecal scores (16%; n = 159), multiple diarrhea severities (10%; n = 95), or the age calves first developed NCD (8%; n = 76). This review characterized substantial heterogeneity among NCD case definitions and diarrhea-related outcomes, which limits interpretation and comparison of studies. Future work is required to develop and validate reporting standards for NCD to optimize knowledge synthesis and support rigorous and ethical calf health research.

Prevalence of susceptibility to Cryptosporidium spp. among dairy calves with different feeding regimens with an emphasis on the feeding of transition milk.

Background and Aim: Colostrum composition and importance for newborn organisms were repeatedly studied. However, the interest in transitional milk usefulness is weak and recommendations concerning transition milk intake are not developed. The aim of this study was to evaluate whether transition milk intake after colostrum consumption affects the chances of calf infection with Cryptosporidium spp. Materials and Methods: We collected data for Cryptosporidium spp. infection from calves (n=425) divided into three groups: The first group - supervised colostrum and transition milk intake; the second group - supervised colostrum and whole milk intake; and the third group - not supervised colostrum and whole milk intake. To detect oocysts of Cryptosporidium spp. in feces, the flotation method was used, and slides were stained using the modified Ziehl-Neelsen method. Generalized linear mixed modeling was conducted to determine whether the explanatory variable - the management of colostrum and transition milk feeding with three categories (three research groups) - was related to the probability of calves incurring infection with Cryptosporidium spp. Results: In the first group, 26.1% of calves were positive for the presence of Cryptosporidium spp. oocysts, in the second - 37.2%, and in the third - 44.1%. Statistical data analysis showed that calves who did not receive transition milk after colostrum consumption had increased chances of having Cryptosporidium spp. (by 1.90-2.47 times on average). The main results showed that the management of colostrum and transition milk feeding is related to Cryptosporidium spp. infection, indicating that both colostrum and transitional milk play a significant role in controlling pathogenic infections. Conclusion: The most effective management of colostrum and transition milk feeding against Cryptosporidium spp. infection is the timely intake of an adequate amount of colostrum followed by transitional milk consumption for at least 2 weeks before weaning from the dam.

Pathogens Detected in 205 German Farms with Porcine Neonatal Diarrhea in 2017.

Neonatal diarrhea (ND) is still a frequently observed problem in modern industrial pig production. ND is predominantly caused by bacterial and viral pathogens. The objective of this study was to give an overview of different pathogens involved in ND in Germany. In 2017, a total number of 555 litters from 205 German pig farms with clinical ND were sampled with pooled fecal samples. All samples were analyzed regarding bacterial pathogens by culture and viral pathogens by polymerase chain reaction (PCR). Isolated strains of Clostridium (C.) perfringens, Escherichia (E.) coli, and C. difficile were further characterized by molecular techniques (e.g., PCR). There were 200 litters (36%), out of 555 sampled litters of 205 farms, which were positive for at least one, while most of them were positive for two or more pathogens. Toxin-producing C. perfringens type A could be detected in 122 farms (59.2%), C. difficile in 116 (56.1%), pathogenic E. coli in 79 (38.6%), and Rotavirus type A in 72 (35%). Among E. coli isolates, enterotoxigenic (8.8%) (F4 fimbriae positive (60.0%)) and necrotoxigenic E. coli (5.3%) were the most frequently detected pathotypes. In conclusion, in most of the farms with porcine ND it turned out to be a disease mainly caused by multiple pathogens, predominantly C. perfringens type A, pathogenic E. coli, and Rotavirus type A. Nevertheless, C. difficile and necrotoxigenic E. coli might be emerging pathogens in ND.

Use of a natural herbal-based feed additive containing isoquinoline alkaloids in newborn calves with cryptosporidiosis.

Cryptosporidium infections are one of the most prevalent causes of diarrhea in calves and considered to be one of the major sources of economic loss in livestock production. A global trend is currently underway, in identifying natural and sustainable alternatives to support animal husbandry and production. Isoquinoline alkaloids are known for their anti-inflammatory, antimicrobial, and antioxidant properties in the promotion of gut health. Thus, an experiment was designed to evaluate the effects of natural, herbal-based feed isoquinoline alkaloids to support calves experimentally inoculated with Cryptosporidium parvum. Twenty-six calves were randomly divided into control (CN) (n = 13) and treatment (SG) (n = 13) groups. The SG group received 5 g of feed additive in every milk feeding from 1 to 21 days of age. The CN group received milk without any additives. All calves were orally inoculated on the third day of life with 1 × 106Cryptosporidium parvum oocysts. The animals were evaluated daily, from 3 to 30 days of age, for the occurrence, duration, and intensity of diarrhea. Calves with a base deficit of ≥ 9 mEq/L were hydrated to aid recovery. The SG calves showed a higher average weight gain between 14 and 21 days of age, without mortality and with reduced intensity and duration of diarrhea. In contrast, calves in the CN group showed more serious acid-base disorders, required more hydration support, and had a mortality rate of 15.4 %. These results showed that calves supplemented with isoquinoline alkaloids had decreased intensity and duration of symptoms, reduced requirement for supportive therapy, and prevented mortality among animals.

Morbidity and mortality in Holstein calves from birth to 145 days of age on a large dairy farm in Brazil / Morbidade e mortalidade em bezerras da raça Holandesa do nascimento aos 145 dias de vida, em uma grande fazenda produtora de leite no Brasil

The present study aimed to investigate the morbidity and mortality associated with the most common diseases during the rearing period of Holstein calves in a high-production dairy farm. The calves (n = 600) were allocated to three groups based on age: G1 (1-7 days, n = 216), G2 (30-40 days, n = 188), and G3 (69-85 days, n = 196). They were evaluated over 60 days at eight different times. The morbidity rates were 47.67% (286/600) for diarrhea, 73.00% (438/600) for bovine respiratory disease (BRD), and 3.83% (23/600) for umbilical inflammation (UI). The mortality rate was 2.33% (14/600) for all the animals studied. The morbidity differed in the groups, with higher rates of diarrhea (78.85%, 166/216) and UI (6.94%, 15/216) in G1, and the highest rate of BRD in G3 (79.59%, 156/196). Diarrhea was associated with the occurrence of BRD in G1 and G2. Even in farms with good management practices, there is high morbidity associated with diarrhea and BRD, with the morbidity rate for diarrhea decreasing and that for BRD increasing as the animals get older. In addition, diarrhea is an important risk factor for BRD, especially in younger animals.(AU)

O presente estudo objetivou investigar a morbidade e a mortalidade associadas às doenças mais comuns no período de criação de bezerras da raça Holandesa, em uma fazenda leiteira de alta produção. As bezerras (n = 600) foram alocadas em três grupos de acordo com a idade: G1 (1-7 dias, n = 216); G2 (30-40 dias, n = 188); G3 (69-85 dias, n = 196). Os animais foram avaliados durante 60 dias, em oito momentos diferentes. A morbidade foi de 47,67% (286/600) para diarreia, 73,00% (438/600) para doença respiratória bovina (DRB) e 3,83% (23/600) para inflamação umbilical (IU). A taxa de mortalidade foi de 2,33% (14/600), para o total de animais avaliados. A morbidade foi diferente entre os grupos, com maior frequência de diarreia (78,85%, 166/216) e IU (6,94%, 15/216) no G1, sendo a maior ocorrência de DRB no G3 (79,59%, 156/196). A diarreia foi associada à DRB no G1 e no G2. Mesmo em fazendas com boas práticas de manejo, há alta morbidade por diarreia e DRB, com a taxa de morbidade por diarreia diminuindo e a de DRB aumentando à medida que os animais envelhecem. Além disso, a diarreia é um importante fator de risco para DRB, especialmente em animais mais jovens.(AU)

Gut health, stress, and immunity in neonatal dairy calves: the host side of host-pathogen interactions.

The cumulative evidence that perinatal events have long-lasting ripple effects through the life of livestock animals should impact future nutritional and management recommendations at the farm level. The implications of fetal programming due to malnutrition, including neonatal survival and lower birth weights, have been characterized, particularly during early and mid-gestation, when placental and early fetal stages are being developed. The accelerated fetal growth during late pregnancy has been known for some time, while the impact of maternal stressors during this time on fetal development and by extent its postnatal repercussions on health and performance are still being defined. Maternal stressors during late pregnancy cannot only influence colostrogenesis but also compromise adequate intestinal development in the fetus, thus, that further limits the newborn's ability to absorb nutrients, bioactive compounds, and immunity (i.e., immunoglobulins, cytokines, and immune cells) from colostrum. These negative effects set the newborn calf to a challenging start in life by compromising passive immunity and intestinal maturation needed to establish a mature postnatal mucosal immune system while needing to digest and absorb nutrients in milk or milk replacer. Besides the dense-nutrient content and immunity in colostrum, it contains bioactive compounds such as growth factors, hormones, and cholesterol as well as molecular signals or instructions [e.g., microRNAs (miRNAs) and long non-coding RNAs (lncRNAs)] transferred from mother to offspring with the aim to influence postnatal gut maturation. The recent change in paradigm regarding prenatal materno-fetal microbiota inoculation and likely the presence of microbiota in the developing fetus intestine needs to be addressed in future research in ruminants. There still much to know on what prenatal or postnatal factors may predispose neonates to become susceptible to enteropathogens (e.g., enterotoxigenic Escherichia coli), causing diarrhea. From the host-side of this host-pathogen interaction, molecular data such as fecal RNA could, over time, help fill those gaps in knowledge. In addition, merging this novel fecal RNA approach with more established microbiome techniques can provide a more holistic picture of an enteropathogenesis and potentially uncover control points that can be addressed through management or nutrition at the farm level to minimize preweaning morbidity and mortality.

Antimicrobial Resistance Profiles and Characterization of Escherichia coli Strains from Cases of Neonatal Diarrhea in Spanish Pig Farms.

Escherichia coli is considered one of the most common agents associated with neonatal diarrhea in piglets. The aim of this work was to characterize the pathogenic and antimicrobial resistance (AMR) profiles of 122 E. coli strains isolated from pigs suffering diarrhea (n = 94) and pigs without diarrhea (n = 28) of 24 farms in Spain. Virulence factors, toxins and AMR (ESBL and colistin) genes and AMR phenotypes of E. coli isolates were analyzed. Low prevalence of pathogenic E. coli strains (26%) was found in both groups. However, ETEC and VTEC strains were more frequently isolated from diarrheic piglets. Irrespectively of diarrhea occurrence, 97.5% of the strains showed a multidrug-resistance (MDR) profile to aminopenicillins, sulfonamides and tetracyclines. It was found that 22% of E. coli was CTX-M+, with CTX-M-14 being the principal allelic variant. Remarkably, 81.5% of CTX-M+ strains were isolated from diarrheic animals and presented an extended MDR profile to aminopenicillins, quinolones and aminoglycosides. Finally, low frequencies of colistin resistance genes mcr-1 (4/122) and mcr-4 (1/122) were found. MDR E. coli strains are circulating in pig farms of Spain, representing a serious threat to animal and public health. More appropriate diagnostic approaches (genetic and AMR phenotypic analysis) should be implemented in animal health to optimize antibiotic treatments.

Congenital chloride diarrhea and Pendred syndrome: case report of siblings with two rare recessive disorders of SLC26 family genes.

BACKGROUND: Congenital chloride diarrhea (CLD; OMIM 214700) is a rare autosomal recessive disorder caused by pathogenic variations in the solute carrier family 26 member A3 (SLC26A3) gene. Without salt substitution, this chronic diarrheal disorder causes severe dehydration and electrolyte disturbances. Homozygous variants in the nearby gene SLC26A4 disrupt anion exchange in the inner ear and the thyroid, causing Pendred syndrome (PDS; OMIM 274600), which is the most frequent form of syndromic deafness. CASE PRESENTATION: We report an unusual co-occurrence of two rare homozygous mutations in both the SLC26A3 and SLC26A4 genes, causing a rare combination of both CLD and PDS in two siblings. Although the clinical pictures were typical, the combined loss of these anion transporters might modulate the risk of renal injury associated with CLD. CONCLUSIONS: Familial presentation of two rare autosomal recessive disorders with loss of function of different SLC26 anion transporters is described. Independent homozygous variants in the SLC26A3 and SLC26A4 genes cause CLD and PDS in siblings, shedding light on co-occurrence of rare recessive traits in the progeny of consanguineous couples.

Prevalence of Virulence Genes and Antimicrobial Resistances in E. coli Associated with Neonatal Diarrhea, Postweaning Diarrhea, and Edema Disease in Pigs from Austria.

Increasing numbers of multi-resistant Escherichia (E.) coli from clinical specimens emphasize the importance of monitoring of their resistance profiles for proper treatment. Furthermore, knowledge on the presence of virulence associated genes in E. coli isolates from European swine stocks is scarce. Consequently, a total of 694 E. coli isolated between 2016 and 2018 from diarrheic piglets of Austrian swine herds were investigated. The isolates were tested for their susceptibility to twelve antibiotics using agar disk diffusion test and for the presence of 22 virulence associated genes via PCR. Overall, 71.9, 67.7, and 49.5% of all isolates were resistant to ampicillin, tetracycline, and trimethoprim-sulfamethoxazole, while resistance levels to gentamicin and fosfomycin were 7.7 and 2.0%, respectively. Resistance frequency to ciprofloxacin was higher than in previous studies. Isolates were more likely to be resistant to ampicillin if they were also resistant to ciprofloxacin. No isolate was resistant to meropenem or amikacin. Virulence genes were detected more frequently in isolates expressing hemolytic activity on blood agar plates. The detection rate of faeG was increased in fimH negative isolates. We assume, that hemolytic activity and absence of fimH could be considered as potential indicators for the virulence of E. coli in piglets.

Prevalence of enteric pathogens in diarrheic and non-diarrheic samples from pig farms with neonatal diarrhea in the North East of Spain.

Diarrhea is one of the major causes of neonatal mortality in pigs. In the present study, 31 pig farms with outbreaks of neonatal diarrhea were investigated in Catalonia (NE Spain) from February 2017 until June 2018. Two hundred and fifteen diarrheic samples from 1 to 7 days old piglets were tested for a panel of enteric pathogens. In 19 of the studied farms additional fecal samples from apparently healthy pen-mates were collected and tested for the same panel of infectious agents. Samples were bacteriologically cultured and tested by PCR for E. coli virulence factors genes, C. perfringens types A and C toxins (Cpα, Cpß, Cpß2) and C. difficile toxins (TcdA, TcdB). Moreover, Rotavirus A (RVA), Rotavirus B (RVB), Rotavirus C (RVC), porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) were also determined by RT-qPCR. More than one pathogen could be detected in all of the outbreaks. Nevertheless, RVA was the only agent that could be statistically correlated with the outcome of diarrhea. For the other viruses and bacteria analyzed significant differences between the diseased pigs and the controls were not found. In spite of this, the individual analysis of each of the studied farms indicated that other agents such as RVB, RVC, toxigenic C. difficile or pathogenic E. coli could play a relevant role in the outbreak of diarrhea. In conclusion, the large diversity of agent combinations and disease situations detected in neonatal diarrhea outbreaks of this study stand for a more personalized diagnosis and management advice at a farm level.

Phenotypic and genotypic characterization of enterotoxigenic Escherichia coli isolated from diarrheic calves in Argentina.

Enterotoxigenic Escherichia coli (ETEC) is the most common and global cause of neonatal calf diarrhea, but there is a little information regarding calf ETEC strains in Argentina. In this study, five ETEC isolates from diarrheic dairy calves (2-10 d old) from Buenos Aires and Cordoba, Argentina were characterized on the basis of virulence gene (VG) pattern, O:H serotyping, hemolytic phenotype, phylogenetic group affiliation, antimicrobial (AM) resistance profile, and presence of integron class 1 and 2. The five isolates were examined by polymerase chain reaction (PCR) for the presence of 18 bovine VGs and showed the following genotypes: F5+/F41+/sta+ (D242), F5+/sta+ (D158), F5+/sta+ (D157), F5+ (D151-9), and F5+/iucD+ (D151-5). These VGs confer pathogenic potential and most of them are associated with the ETEC pathotype. The five isolates showed a non-hemolytic phenotype, belonged to five different serotypes: O101:H-, O141:H-, O60:H-, ONT:H10, and ONT:H-, and were assigned to the phylogenetic group A by the quadruplex Clermont PCR method. The AM resistance of the three isolates D242, D157, and D151-5 was determined by agar disk diffusion method for 24 AMs and they exhibited a multi-resistance phenotype (resistance to four different AM classes: Cephalosporins, Penicillins, Macrolides, and Ansamycins). In addition, class 1 integrons were found in the isolate D151-5 containing the dfrA17-aadA5 gene cassette and in the bovine ETEC reference strain FV10191 containing the dfrA1-aadA1 gene cassette. The present study revealed for the first time the occurrence of multi-resistant ETEC associated with neonatal diarrhea in dairy calves in Argentina. This finding may be used for diagnostic and therapeutic purposes.

Long-term use of yeast fermentation products in comparison to halofuginone for the control of cryptosporidiosis in neonatal calves.

The objective of this study was to compare the effect of non-GMO Saccharomyces cerevisiae fermentation products (SCFP) with that of a halofuginone treatment against Cryptosporidium parvum infection in pre-weaned calves on a commercial dairy farm. A total of 123 neonatal female calves, housed in individual hutches, were enrolled sequentially based on date of birth in 41 blocks of 3 animals each. Calves within each block were allocated to one of 3 treatments: remaining untreated, fed with SCFP (Diamond V SmartCare® at 1 g/d in milk and NutriTek® at 5 g/d in starter grain) for the first 63 days of life, or treated with halofuginone (0.1 mg/kg/d) for the first 7 days of life. Fecal samples collected on days 4-21 post-partum were examined for both Cryptosporidium oocysts and coproantigen. The presence and intensity of diarrhea were monitored by scoring daily for the first 4 weeks of life. Calves were weighed at 0, 21, 42 and 63 days of age. Almost all calves were Cryptosporidium-positive at least once during the study. Halofuginone significantly reduced the number of Cryptosporidium-positive fecal samples as compared to the two other groups. Based on the coproantigen scores, both halofuginone and SCFP feeding significantly reduced the intensity of Cryptosporidium infection as compared to the untreated group. Diarrhea was recorded in almost all calves at least once. Neither the proportion of diarrheic calves nor the intensity and duration of diarrhea differed among the 3 treatment groups significantly. The mean daily weight gain during the first 3 weeks of life was significantly lower in halofuginone treated calves than in both other groups; however, at the end of the study period the total weight gain did not significantly differ among the 3 treatment groups. In conclusion, the clinical results and weight gains of pre-weaning supplementation with the SCFP were neither better nor worse than the 7-day halofuginone treatment suggesting that the SCFP feeding may be from the clinical point of view a natural alternative measure, instead of halofuginone treatment, in bovine cryptosporidiosis.

Longitudinal surveillance of rotavirus A genotypes circulating in a high milk yield dairy cattle herd after the introduction of a rotavirus vaccine.

Worldwide, neonatal diarrhea is one of the most important health issues affecting dairy calves, and rotavirus A (RVA) is one of its primary causes. Among the measures to mitigate the risk of diarrhea outbreaks, cow vaccination stands out as one of the most important. However, the immune pressure resulting from routine vaccination may be able to select specific G and P genotypes in RVA field strains. This study aimed to determine the frequency and intensity of neonatal diarrhea and the incidence of RVA and attempted to monitor the G and P genotypes present in the RVA strains circulating in a high milk yield cattle herd vaccinated with RVA G6P[5] strain. Fecal samples (n = 1220) from 122 Holstein heifer calves between 0-30 days old that were born from RVA-vaccinated cows were collected at 10 different time points, regardless of the presence or absence of diarrhea. The presence of RVA in fecal samples was determined by the polyacrylamide gel electrophoresis (PAGE) technique and confirmed by reverse transcription polymerase chain reaction (RT-PCR). G and P amplicons from 10 RVA-positive fecal samples from calves of different ages and collections were subjected to nucleotide sequencing. The proportion of the calves and fecal samples that were positive for RVA were 62.3% (76/122) and 8.1% (99/1220), respectively. Using sequence analysis, all 10 RVA field strains presented genotype G10P[11]. The protection of G6P[5] vaccination is clear, as this genotype was not detected in this study, and it is known that vaccination against RVA reduces the incidence of diarrhea independent of genotype involved. This result demonstrates the importance of epidemiological monitoring of RVA genotypes circulating in vaccinated dairy cattle herds to the early detection of new potential pathogenic RVA strains.

Detection of endotoxin in plasma of hospitalized diarrheic calves.

OBJECTIVES: To investigate whether lipopolysaccharide (LPS) is present in plasma of calves with naturally occurring diarrhea. The second objective was to determine whether plasma [LPS] correlates with clinical, hematological, biochemical, and acid-base variables, and whether [LPS] differs between surviving and nonsurviving diarrheic calves. DESIGN: Prospective observational study (January 2012-May 2014). SETTING: Veterinary teaching hospital. ANIMALS: Thirty-four calves <28 days old admitted for diagnosis and treatment of diarrhea and 30 healthy control calves. MEASUREMENTS AND MAIN RESULTS: Admission demographics, physical examination, blood gas, biochemistry analysis, and outcome data were recorded. Plasma concentration of LPS was determined using a bovine LPS ELISA assay. Plasma [LPS] was detected in both healthy and diarrheic calves. Plasma [LPS] was significantly higher in diarrheic than healthy calves (median: 0.99 ng/mL; Interquartile range (IQR): 0.068, vs 0.88 ng/mL; 0.065 ng/mL, respectively; P < 0.001). Plasma [LPS] was higher in nonsurviving (1.04 ng/mL; 0.07 ng/mL) than in surviving calves (0.98 ng/mL; 0.022 ng/mL; P < 0.001). Plasma [LPS] was higher in beef (1.07 ng/mL; 0.182 ng/mL) than in dairy diarrheic calves (0.99 ng/mL; 0.022 ng/mL; P < 0.001). In diarrheic calves, plasma [LPS] correlated with [l-lactate] (r2 = 0.496; P = 0.002); hypoglycemia (r2 = -0.453; P = 0.007); increased unmeasured strong ions (r2 = 0.332; P = 0.050), [Mg2+ ] (r2 = 0.475; P = 0.004), and [phosphate] (r2 = 0.468; P = 0.005), and increased aspartate aminotransferase activity (r2 = 0.348; P = 0.003). CONCLUSIONS: This study highlights a potential role of LPS in the pathogenesis of metabolic derangements such as hyperlactatemia, hypoglycemia, and increased concentration of unmeasured strong anions in diarrheic calves. Further investigation evaluating the effect of LPS on l-lactate and glucose metabolism in diarrheic calves is warranted.

Effect of prebiotic supplementation with stabilized rice bran in milk of pre-weaned organic Holstein calves.

BACKGROUND: The first month of life possess significant challenges for dairy calves due to high susceptibility to digestive diseases. The objective of this study was to evaluate the effect of prebiotic supplementation with stabilized rice bran (SRB) in milk on health, immunity, and performance of pre-weaned organic dairy calves. Holstein heifer calves (n = 90) were enrolled at 6 ± 1 days old and monitored for 28 days, from July to August 2017. Calves were randomly assigned to a control (CTR; n = 45) or a treatment group (SRB; n = 45). The CTR group received milk alone and the SRB group received 120 g of SRB per day in milk to achieve a 10% w/w dose of the total calories. Daily health evaluations were conducted to score health status and disease severity (healthy, slightly affected, moderately or severely sick) of calves, through integrated assessment of diarrhea, dehydration, attitude, and milk intake. Body weights and fecal IgA quantification were completed on the first and last day of the study. RESULTS: Overall, weight gain and fecal IgA concentrations were not affected by the dietary addition of SRB. The total number of calf-days classified as healthy or sick were not different between treatment groups. Similarly, the number of calf-days categorized as slightly affected, moderately sick, or severely sick did not differ between treatment groups. Time to event analyses indicated a tendency for a treatment effect in the time to the first moderate case of diarrhea (P = 0.08), as well as in the time to recovery from diarrhea (P = 0.052), favoring control calves. CONCLUSIONS: These results indicated that the dietary addition of SRB in milk did not have an effect in health, immunity or performance of pre-weaned dairy calves.