Comparison of the fecal microbiota of adult healthy dogs fed a plant-based (vegan) or an animal-based diet.

Purpose: Pet guardians are increasingly seeking vegan dog foods. However, research on the impact of these diets on gastrointestinal (GI) physiology and health is limited. In humans, vegan diets modify the GI microbiota, increasing beneficial digestive microorganisms. This study aimed to examine the canine fecal microbiota in response to a vegan diet compared to an animal-based diet. Methods: Sixty-one client-owned healthy adult dogs completed a randomized, double-blinded longitudinal study. Dogs were randomly assigned into two groups that were fed either a commercial extruded animal-based diet (MEAT, n = 30) or an experimental extruded vegan diet (PLANT, n = 31) for 12 weeks. Fecal collections occurred at the start of the experimental period and after 3 months of exclusively feeding either diet. Bacterial DNA was extracted from the feces, and the V4 region of the 16S rRNA gene was amplified using PCR and sequenced on Illumina MiSeq. Beta-diversity was measured using Jaccard and Bray-Curtis distances, and the PERMANOVA was used to assess for differences in fecal microbiota within and between groups. Alpha-diversity indices for richness, evenness, and diversity, as well as relative abundance, were calculated and compared between groups. Results: Beta-diversity differences occurred between diet groups at exit time-point with differences on Bray-Curtis distances at the family and genus levels (p = 0.007 and p = 0.001, respectively), and for the Jaccard distance at the family and genus level (p = 0.006 and p = 0.011, respectively). Significant differences in alpha-diversity occurred when comparing the PLANT to the MEAT group at the exit time-point with the PLANT group having a lower evenness (p = 0.012), but no significant differences in richness (p = 0.188), or diversity (p = 0.06). At exit-timepoint, compared to the MEAT group, the relative abundance of Fusobacterium, Bacteroides, and Campylobacter was lower in the PLANT group. The relative abundance of Fusobacterium decreased over time in the PLANT group, while no change was observed in the MEAT group. Conclusion: These results indicate that vegan diets may change the canine gut microbiota. Future studies are warranted to confirm our results and determine long-term effects of vegan diets on the canine gut microbiome.

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 the effects of transport duration on the fecal microbial communities of surplus dairy calves.

Transportation significantly impacts the health and welfare of surplus dairy calves largely due to the various stressors and pathogen exposures encountered during the process. Concurrently, the animal's microbiome is known to correlate with its health status, with stress-induced alterations in the microbiota potentially precipitating various diseases. This study aimed to compare the impacts of transportation durations of 6, 12, or 16 h on the fecal microbiota in young surplus dairy calves. We used a randomized controlled design, where surplus dairy calves aged 1-19 d old from 5 commercial dairy farms in Ontario were allocated into one of 3 (6, 12, and 16 h of continuous transportation) transportation groups. Health assessments were conducted before, immediately after, and for 2 weeks following transportation. Fecal samples were collected before, immediately after, and at 24- and 72 h post-transportation and subjected to 16S rRNA sequencing. Alpha diversity metrics showed no significant differences between the 3 transportation groups at any of the sampling time points. Although ß diversity metrics revealed no clustering by transportation groups, they indicated significant differences across sampling time points within each group. The overall analysis revealed a total of 22 phyla and 353 genera, with Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria, and Fusobacteria being the most abundant phyla. Bacteroides, Escherichia/Shigella, Lactobacillus, Collinsella, and Bifidobacterium were the most abundant genera. The reduction in Fusobacteria abundance before and after transport was significantly larger in the 16-h transportation group when compared with the 6-h transportation group. We also identified several genus-level and ASV-level taxa that displayed significant differences in their abundances across various transportation groups, observed at all sampling time points investigated. In conclusion, this research identifies microbiota changes due to varying transportation durations in surplus dairy calves providing a broad understanding of the microbial shifts in surplus dairy calves' post-transportation across varying durations. While these variations may not directly correlate with overall calf health or indicate dysbiosis, these results emphasize the importance of further investigating transportation practices to enhance calf health and well-being. Further studies are warranted to elucidate the relationship between microbiota and calf health.

Impact of fecal sample preservation and handling techniques on the canine fecal microbiota profile.

Canine fecal microbiota profiling provides insight into host health and disease. Standardization of methods for fecal sample storage for microbiomics is currently inconclusive, however. This study investigated the effects of homogenization, the preservative RNAlater, room temperature exposure duration, and short-term storage in the fridge prior to freezing on the canine fecal microbiota profile. Within 15 minutes after voiding, samples were left non-homogenized or homogenized and aliquoted, then kept at room temperature (20-22°C) for 0.5, 4, 8, or 24 hours. Homogenized aliquots then had RNAlater added or not. Following room temperature exposure, all aliquots were stored in the fridge (4°C) for 24 hours prior to storing in the freezer (-20°C), or stored directly in the freezer. DNA extraction, PCR amplification, then sequencing were completed on all samples. Alpha diversity (diversity, evenness, and richness), and beta diversity (community membership and structure), and relative abundances of bacterial genera were compared between treatments. Homogenization and RNAlater minimized changes in the microbial communities over time, although minor changes in relative abundances occurred. Non-homogenized samples had more inter-sample variability and greater changes in beta diversity than homogenized samples. Storage of canine fecal samples in the fridge for 24 h prior to storage in the freezer had little effect on the fecal microbiota profile. Our findings suggest that if immediate analysis of fecal samples is not possible, samples should at least be homogenized to preserve the existing microbiota profile.

Survival rates and factors associated with survival and laminitis of horses with acute diarrhoea admitted to referral institutions.

BACKGROUND: Clinicopathological findings and their association with the outcome and development of laminitis in horses with acute diarrhoea has not been investigated in a multicentre study across different geographic regions. OBJECTIVES: Describe and compare clinicopathologic findings of diarrhoeic horses between different geographic regions, survival rates and factors associated with non-survival and laminitis. STUDY DESIGN: Multicentre retrospective case series. METHODS: Information from horses with acute diarrhoea presenting to participating institutions between 2016 and 2020 was collected, and clinicopathological data were compared between surviving and non-surviving horses and horses that did and did not develop laminitis. Survival rates and seasonal and geographic differences were also investigated. RESULTS: One thousand four hundred thirty-eight horses from 26 participating institutions from 4 continents were included; 76% survived to discharge with no differences identified between geographic regions. The survival proportion of horses with SIRS and creatinine concentrations > 159 µmol/L was 55% (154/279) compared with 81% (358/437) for those with SIRS and creatinine concentrations < 159 µmol/L (p < 0.001). The survival proportion of horses with SIRS that had an L-lactate concentration > 2.8 mmol/L was 59% (175/298) compared with 81% (240/296) in horses with SIRS and L-lactate concentration < 2.8 mmol/L (p < 0.001). The proportion of horses that developed laminitis was lower in Europe (4%, 19/479) compared with North America (8%, 52/619), Australia (8%, 12/138) and Latin America (11%, 16/146) (p < 0.05). More horses developed laminitis in the summer (46%, 39/85) compared with winter (18%, 15/85), spring (18%, 15/85) and fall (19%, 16/85) (p < 0.01). Horses with laminitis had greater odds of non-survival than those without laminitis (OR: 3.73, 95% CI: 2.47-5.65). MAIN LIMITATIONS: Not all variables were available for all horses due to the retrospective nature. CONCLUSIONS: Clinicopathological findings in horses with acute diarrhoea and their association with survival are similar across geographic regions. However, developing laminitis secondary to diarrhoea is less common in Europe. In addition, factors associated with non-survival were indicative of disease severity and subsequent cardiovascular compromise.

CONTEXTO: Achados clínico-patológicos e suas associações com o sobrevivência e desenvolvimento de laminite em cavalos com diarreia aguda não foram investigados em um estudo multicêntrico envolvendo diferentes regiões geográficas. OBJETIVOS: Descrever e comparar achados clínico-patológicos de cavalos com diarreia em diferentes regiões geográficas, taxa de sobrevivência e fatores associados com mortalidade e laminite. DELINEAMENTO DO ESTUDO: Estudo multicêntrico retrospectivo de série de casos. METODOLOGIA: Informação sobre equinos com diarreia aguda apresentados às instituições participantes entre 2016 e 2020 foram coletados, e dados clínico-patológicos foram comparados entre sobreviventes e não-sobreviventes, e cavalos que desenvolveram ou não laminite. Taxa de sobrevivência, e diferenças sazonais e geográficas também foram investigadas. RESULTADOS: 1438 cavalos de 26 instituições participantes de 4 continentes foram incluídos; 76% sobreviveram e receberam alta e nenhuma diferença foi observada entre as diferentes regiões geográficas. A proporção de cavalos que sobreviveram com SIRS e concentração de creatinina > 1.8 mg/dL foi 55% (154/279) comparado com 81% (358/437) dos cavalos com SIRS e concentração de creatinina < 1.8 mg/dL (p < 0.001). A proporção de cavalos com SIRS que tinham concentração de L-lactato > 2.8 mmol que sobreviveram foi 59% (175/298) comparado com 81% (240/296) dos cavalos com SIRS e concentração de L-lactato < 2.8 mmol/L (p < 0.001). A proporção de cavalos que desenvolveram laminite foi menor na Europa (4%, 19/479) comparado com a América do Norte (8%, 52/619), Austrália (8%, 12/138) e América Latina (11% 16/146) (p < 0.05). Mais cavalos desenvolveram laminite no verão (46%, 39/8) comparado com inverno (18%, 15/85), primavera (18%, 15/85) e outono (19%, 16/85) (p < 0.01). Cavalos com laminite tiveram chances maior de não sobreviver do que aqueles que não desenvolveram laminite (OR: 3.73, 95% CI: 2.47 a 5.65). PRINCIPAIS LIMITAÇÕES: Algumas variáveis não estavam disponíveis para alguns cavalos devido à natureza retrospectiva deste estudo. CONCLUSÕES: Achados clínico-patológicos em equinos com diarreia aguda e sua associação com sobrevivência é similar entre as diferentes regiões geográficas. Contudo, o desenvolvimento de laminite secundário à diarreia é menos comum na Europa. Além disso, fatores associados com não-sobrevivência foram indicativos de severidade da doença e subsequente comprometimento cardiovascular.

Diagnostic approaches, aetiological agents and their associations with short-term survival and laminitis in horses with acute diarrhoea admitted to referral institutions.

BACKGROUND: An international description of the diagnostic approaches used in different institutions to diagnose acute equine diarrhoea and the pathogens detected is lacking. OBJECTIVES: To describe the diagnostic approach, aetiological agents, outcome, and development of laminitis for diarrhoeic horses worldwide. STUDY DESIGN: Multicentre retrospective case series. METHODS: Information from horses with acute diarrhoea presenting to participating institutions between 2016 and 2020, including diagnostic approaches, pathogens detected and their associations with outcomes, were compared between institutions or geographic regions. RESULTS: One thousand four hundred and thirty-eight horses from 26 participating institutions from 4 continents were included. Overall, aetiological testing was limited (44% for Salmonella spp., 42% for Neorickettsia risticii [only North America], 40% for Clostridiodes difficile, and 29% for ECoV); however, 13% (81/633) of horses tested positive for Salmonella, 13% (35/262) for N. risticii, 9% (37/422) for ECoV, and 5% (27/578) for C. difficile. C. difficile positive cases had greater odds of non-survival than horses negative for C. difficile (OR: 2.69, 95%CI: 1.23-5.91). In addition, horses that were positive for N. risticii had greater odds of developing laminitis than negative horses (OR: 2.76, 95%CI: 1.12-6.81; p = 0.029). MAIN LIMITATIONS: Due to the study's retrospective nature, there are missing data. CONCLUSIONS: This study highlighted limited diagnostic investigations in cases of acute equine diarrhoea. Detection rates of pathogens are similar to previous reports. Non-survival and development of laminitis are related to certain detected pathogens.

Association of the type of metabolic acidosis and non-survival of horses with colitis.

Background: Metabolic acidosis (MA) is the most common acid-base disorder reported in horses with colitis but its association with survival is yet to be determined. Objective: Investigate the types of MA in horses with colitis to determine effects of various anions on fatality rates. Animals and procedures: We studied 158 horses with colitis. Horses were classified into 4 groups depending on the anion contributing to MA: i) no MA, ii) lactic acidosis (LA), iii) unmeasured strong ion (USI) acidosis, and iv) hyperchloremic acidosis (HA). Results: Sixty percent (95/158) of horses had no MA, 22% (34/158) had LA, 12% (19/158) had HA, and 6% (10/158) had USI acidosis. The fatality rate of horses without MA was 20% (20/95), whereas the rates for those with LA, USI, and HA were 53% (18/34), 30% (3/10), and 16% (3/19), respectively. Horses with LA were more likely to die or be euthanized than horses without MA (OR: 4.2, 95% CI: 1.83 to 9.72, P < 0.001) and HA (OR: 5.9, 95% CI: 1.47 to 24.4, P < 0.01). Conclusion and clinical relevance: Lactic acidosis was the most common type of MA in horses with colitis, and it was associated with non-survival.

Association du type d'acidose métabolique et de non-survie des chevaux atteints de colite. Historique: L'acidose métabolique (AM) est le trouble acido-basique le plus fréquemment signalé chez les chevaux atteints de colite, mais son association avec la survie reste à déterminer. Objectif: Étudier les types d'AM chez les chevaux atteints de colite pour déterminer les effets de divers anions sur les taux de mortalité. Animaux et procédures: Nous avons étudié 158 chevaux atteints de colite. Les chevaux ont été classés en 4 groupes en fonction de l'anion contribuant à l'AM : i) pas d'AM, ii) acidose lactique (LA), iii) acidose à ions forts non mesurés (USI) et iv) acidose hyperchlorémique (HA). Résultats: Soixante pour cent (95/158) des chevaux n'avaient pas d'AM, 22 % (34/158) avaient une LA, 12 % (19/158) avaient une HA et 6 % (10/158) avaient une acidose USI. Le taux de mortalité des chevaux sans AM était de 20 % (20/95), tandis que les taux de ceux avec LA, USI et HA étaient de 53 % (18/34), 30 % (3/10) et 16 % (3/19), respectivement. Les chevaux atteints de LA étaient plus susceptibles de mourir ou d'être euthanasiés que les chevaux sans AM (OR : 4,2, IC à 95 % : 1,83 à 9,72, P < 0,001) et HA (OR : 5,9, IC à 95 % : 1,47 à 24,4, P < 0,01). Conclusion et pertinence clinique: L'acidose lactique était le type d'AM le plus courant chez les chevaux atteints de colite et elle était associée à la non-survie.(Traduit par Dr Serge Messier).

Rostral spread of lumbosacral epidural volumes of dye and contrast medium calculated using body weight or length of the vertebral column in dog cadavers.

The objective of this study was to compare the rostral spread of lumbosacral epidural volumes of a mixture of dye and contrast medium, calculated using body weight (BW) or vertebral column length (LE), in 22 dog cadavers. The dogs weighed 4.6 to 52.0 kg. Dogs were paired within a < 10% difference for BW and LE and with the same body condition score (BCS). Pairs of dogs were injected while in sternal recumbency through an epidural catheter with a volume mixture of iopamidol and dye, calculated based on BW: 0.2 mL/kg in one of the cadavers and based on LE: 0.05 mL/cm (< 50 cm), 0.07 mL/cm (50 to < 70 cm), 0.08 mL/cm (70 to < 80 cm), and 0.11 mL/cm (≥ 80 cm) in the other cadaver. The extent of rostral spread was determined using computed tomography for iopamidol and anatomical dissection for dye. Comparisons for dye and iopamidol within each dog, and for BW and LE within matched pairs, were completed with mixed linear models (P < 0.05). The number of vertebrae reached by dye was greater than the number reached by iopamidol in both BW and LE, but the rostral spread was not significantly different between BW and LE for all pairs. In conclusion, dye tends to spread further than iopamidol and therefore, these two methods should not be considered interchangeable when used in research studies.

L'objectif de cette étude était de comparer l'étalement rostral des volumes épiduraux lombo-sacrés d'un mélange de colorant et de produit de contraste, calculé en utilisant le poids corporel (PC) ou la longueur de la colonne vertébrale (LE), chez 22 cadavres de chiens. Les chiens pesaient de 4,6 à 52,0 kg. Les chiens ont été appariés avec une différence < 10 % pour PC et LE et avec le même score d'état corporel (PCS). Des paires de chiens ont reçu une injection alors qu'ils étaient en décubitus sternal à travers un cathéter péridural avec un mélange volumique d'iopamidol et de colorant, calculé sur la base du poids corporel : 0,2 mL/kg dans l'un des cadavres et sur la base de la LE : 0,05 mL/cm (< 50 cm), 0,07 mL/cm (50 à < 70 cm), 0,08 mL/cm (70 à < 80 cm) et 0,11 mL/cm (≥ 80 cm) dans l'autre cadavre. L'étendue de la propagation rostrale a été déterminée par tomodensitométrie pour l'iopamidol et dissection anatomique pour le colorant. Les comparaisons pour le colorant et l'iopamidol au sein de chaque chien, et pour PC et LE au sein de paires appariées, ont été complétées avec des modèles linéaires mixtes (P < 0,05). Le nombre de vertèbres atteintes par le colorant était supérieur au nombre atteint par l'iopamidol dans PC et LE, mais la propagation rostrale n'était pas significativement différente entre PC et LE pour toutes les paires. En conclusion, le colorant a tendance à se propager plus loin que l'iopamidol et, par conséquent, ces deux méthodes ne doivent pas être considérées comme interchangeables lorsqu'elles sont utilisées dans des études de recherche.(Traduit par Docteur Serge Messier).

Comparison of rostral spread of lumbosacral epidural volume calculated by body weight or length of the vertebral column in small-sized anesthetized dogs.

The first objective of this prospective, randomized, crossover experimental trial was to compare the rostral spread of lumbosacral epidural volume calculated by body weight (BW) or vertebral column length (LE) in 6 small, isoflurane-anesthetized female beagle dogs (BW: 7.5 to 10.2 kg; LE measured from the occipital crest to the sacrococcygeal space: 46 to 56 cm). The second objective was to assess the response to a noxious stimulus once the dogs recovered from anesthesia and to determine the effects of the injection on cardiopulmonary variables. While in sternal position, dogs were injected through an epidural catheter with a volume mixture of bupivacaine 0.25% and iopamidol 15% based on BW: 0.2 mL/kg or LE: 0.05 mL/cm (< 50 cm) or 0.07 mL/cm (50 to < 70 cm). Rostral spread was determined by counting the number of vertebrae reached by iopamidol using computed tomography. After anesthesia, cardiopulmonary variables, motor function, and responses to nociceptive stimuli were evaluated. Comparisons were completed with mixed linear models and 2-way analysis of variance (ANOVA) (P < 0.05). The volume of injectate (3.29 ± 0.74 versus 1.81 ± 0.21 mL; mean ± SD) and the number of vertebrae (22 ± 2 versus 19 ± 2 vertebrae) reached by iopamidol were significantly greater for LE than for BW. Response to nociception, time to return of pain sensation, motor function, and cardiopulmonary variables were similar between groups. In conclusion, dosing based on LE resulted in larger rostral spread than when based on BW in dogs of small size.

Le premier objectif de cet essai expérimental croisé prospectif randomisé était de comparer la propagation rostrale du volume épidural lombo-sacré calculé en fonction du poids corporel (PC) ou de la longueur de la colonne vertébrale (LE) chez 6 petites chiennes beagle anesthésiées à l'isoflurane (PC : 7,5 à 10,2 kg; LE mesuré de la crête occipitale à l'espace sacro-coccygien : 46 à 56 cm). Le deuxième objectif était d'évaluer la réponse à un stimulus nocif une fois que les chiens se sont remis de l'anesthésie et de déterminer les effets de l'injection sur les variables cardiopulmonaires. En position sternale, les chiens ont reçu une injection via un cathéter péridural d'un mélange volumique de bupivacaïne à 0,25 % et d'iopamidol à 15 % basé sur le poids corporel : 0,2 mL/kg ou LE : 0,05 mL/cm (< 50 cm) ou 0,07 mL/cm (50 à < 70 cm). La propagation rostrale a été déterminée en comptant le nombre de vertèbres atteintes par l'iopamidol en utilisant la tomodensitométrie. Après l'anesthésie, les variables cardiopulmonaires, la fonction motrice et les réponses aux stimuli nociceptifs ont été évaluées. Les comparaisons ont été complétées avec des modèles linéaires mixtes et une analyse de variance à 2 facteurs (ANOVA) (P < 0,05). Le volume d'injectat (3,29 ± 0,74 versus 1,81 ± 0,21 mL; moyenne ± SD) et le nombre de vertèbres (22 ± 2 versus 19 ± 2 vertèbres) atteints par l'iopamidol étaient significativement plus élevés pour LE que pour BW. La réponse à la nociception, le temps de retour de la sensation de douleur, la fonction motrice et les variables cardiopulmonaires étaient similaires entre les groupes. En conclusion, le dosage basé sur LE a entraîné une plus grande propagation rostrale que lorsqu'il était basé sur BW chez les chiens de petite taille.(Traduit par Docteur Serge Messier).

Nontuberculous Mycobacteria in Horses: A Narrative Review.

Nontuberculous mycobacteria (NTM) infections are increasing in human and veterinary medicine. Although horses were initially thought to be resistant to NTM infection, reports of horses suffering from gastrointestinal, respiratory, and reproductive diseases associated with NTM have increased in the last few decades. The aim of this literature review is to summarize the mycobacteria species found in horses, describe clinical manifestations, diagnostic and treatment approaches, and public health concerns of NTM infection in horses. Clinical manifestations of NTM in horses include pulmonary disease, lymphadenitis, soft tissue, bone infections, and disseminated disease. NTM are also linked to granulomatous enteritis, placentitis, and abortions. Currently, diagnostic methods for NTM are limited and include acid-fast microscopy, bacterial cultures, species-specific PCR assays, and gene sequencing. In humans, NTM treatment guidelines are available, but their application appears inadequate and inconsistent. In horses, treatment guidelines for NTM infections are not available. NTM are a serious public health threat as 70% of people with untreated acquired immunodeficiency syndrome (AIDS) have a chronic pulmonary disease caused by NTM. Thus, it is essential that we gain a better understanding of NTM infections in horses and their zoonotic potential.

The effects of ambient temperature exposure on feline fecal metabolome.

Introduction: The fecal metabolome provides insight into overall gastrointestinal and microbial health. Methods for fecal sample storage in metabolomics research vary, however, making comparisons within current literature difficult. This study investigated the effect of ambient temperature exposure on microbial-derived metabolites of feline fecal samples. Methods: Fecal samples were collected from 11 healthy cats from a local boarding facility. Samples were manually homogenized and aliquoted. The first aliquot was frozen at -80°C within 1 hour of defecation, and remaining samples were exposed to ambient temperature for 2, 4, 6, 8, 12, and 24 h prior to freezing at -80°C. Fecal metabolites were quantified using 1H NMR spectroscopy. Fifty metabolites were grouped into six categories (27 amino acids, 8 fatty acids, 5 sugars, 3 alcohols, 2 nitrogenous bases, 5 miscellaneous). Results: Concentrations of 20 out of 50 metabolites significantly differed due to ambient temperature exposure (7 amino acids, 6 fatty acids, 2 alcohols, 1 nitrogenous base, 4 miscellaneous). The earliest detected changes occurred 6 h post-defecation for cadaverine and fumaric acid. Discussion: This study shows ambient temperature exposure alters the composition of the feline fecal metabolome, but short-term (up to 4 h) exposure prior to storage in the freezer seems to be acceptable.

Effect of time of sample collection after onset of diarrhea on fecal microbiota composition of calves.

BACKGROUND: The effect of time of sample collection after onset of diarrhea on the fecal microbiota composition of calves is unknown. OBJECTIVE: Compare the fecal microbiota of calves with diarrhea onset on the day of sampling (D <24h), and calves having had diarrhea for >24 to 48 hours (D 24-48h). ANIMALS: Thirty-one diarrheic calves (20 D <24h and 11 D 24-48h), 3 to 7 days of age. METHODS: Cross-sectional study. Diarrhea was defined as a calf with loose feces or watery feces. Assessment of the fecal microbiota was performed by sequencing of 16S ribosomal RNA gene amplicons. RESULTS: Richness and diversity were not statistically different between D <24h and D 24-48h (P > .05), but bacterial membership and structure differed significantly (AMOVA, P < .001 for both comparisons). Linear discriminant analysis effect size (LefSe) showed an enrichment of Faecalibacterium, Phocaeicola, Lachnospiracea, and Lactobacillus in the feces of D <24h calves, whereas Escherichia/Shigella, Ligilactobacillus, Clostridium_Sensu_Stricto, Clostridium_Incerta_Sedis, and Enterococcus were enriched in the D 24-48h calves. CONCLUSION AND CLINICAL IMPORTANCE: Rapid changes in fecal microbiota occur during the first 48 hours of diarrhea with an enrichment of lactic acid-producing bacteria in D <24h followed by an enrichment in Escherichia/Shigella and Clostridium spp. in D 24-48h. The time from diarrhea onset to sampling appears to affect the bacterial composition. Researchers should standardize times for fecal collection based on the time of diarrhea.

Postmortem examination of equids >1-year-old with enterotyphlocolitis in Ontario: a retrospective descriptive study.

Equine enterotyphlocolitis is an inflammatory process of the intestinal tract of horses that is associated with multiple etiologic agents and risk factors. Most clinical cases do not have an etiologic diagnosis. We describe here the pathogens detected and the histologic lesions found in horses with enterotyphlocolitis in Ontario that were submitted for postmortem examination, 2007-2019. We reviewed the medical records of 208 horses that fulfilled inclusion criteria. Cultures were positive in 67 of 208 (32%) equids for Clostridium perfringens, in 16 of 208 (8%) for Clostridioides difficile, and in 14 of 208 (7%) for Salmonella spp.; 6 of 208 (3%) were positive for Neorickettsia risticii by PCR assay. One horse was positive in a Rhodococcus equi PCR assay. All horses tested by PCR assay for equine coronavirus and Lawsonia intracellularis were negative. The histologic lesions were characterized as follows: 6 of 208 (3%) enteritis, 5 of 208 (2%) typhlitis, 104 of 208 (50%) colitis, 37 of 208 (18%) enterocolitis, 45 of 208 (22%) typhlocolitis, and 11 of 208 (5%) enterotyphlocolitis. We strongly recommend standardized testing of diarrheic horses during and/or after postmortem examination, as well as standardized reporting of histologic lesions in enterotyphlocolitis cases.

Host innate immune responses and microbiome profile of neonatal calves challenged with Cryptosporidium parvum and the effect of bovine colostrum supplementation.

Introduction: Calves are highly susceptible to gastrointestinal infection with Cryptosporidium parvum (C. parvum), which can result in watery diarrhea and eventually death or impaired development. With little to no effective therapeutics, understanding the host's microbiota and pathogen interaction at the mucosal immune system has been critical to identify and test novel control strategies. Methods: Herein, we used an experimental model of C. parvum challenge in neonatal calves to describe the clinical signs and histological and proteomic profiling of the mucosal innate immunity and microbiota shifts by metagenomics in the ileum and colon during cryptosporidiosis. Also, we investigated the impact of supplemental colostrum feeding on C. parvum infection. Results: We showed that C. parvum challenged calves experienced clinical signs including pyrexia and diarrhea 5 days post challenge. These calves showed ulcerative neutrophil ileitis with a proteomic signature driven by inflammatory effectors, including reactive oxygen species and myeloperoxidases. Colitis was also noticed with an aggravated mucin barrier depletion and incompletely filled goblet cells. The C. parvum challenged calves also displayed a pronounced dysbiosis with a high prevalence of Clostridium species (spp.) and number of exotoxins, adherence factors, and secretion systems related to Clostridium spp. and other enteropathogens, including Campylobacter spp., Escherichia sp., Shigella spp., and Listeria spp. Daily supplementation with a high-quality bovine colostrum product mitigated some of the clinical signs and modulated the gut immune response and concomitant microbiota to a pattern more similar to that of healthy unchallenged calves. Discussion: C. parvum infection in neonatal calves provoked severe diarrheic neutrophilic enterocolitis, perhaps augmented due to the lack of fully developed innate gut defenses. Colostrum supplementation showed limited effect mitigating diarrhea but demonstrated some clinical alleviation and specific modulatory influence on host gut immune responses and concomitant microbiota.

Direct and culture-enriched 16S rRNA sequencing of cecal content of healthy horses and horses with typhlocolitis.

Next generation sequencing has demonstrated that alpha diversity of the fecal microbiota is significantly altered in horses with typhlocolitis. The objective of this study was to evaluate the bacterial composition of the cecum content of horses with and without typhlocolitis through direct and culture-enriched 16S gene sequencing of six healthy horses and six horses with acute typhlocolitis; a case-control study design. Cecal content was collected after euthanasia. An aliquot was used for direct 16S gene sequencing. Another was serially diluted with brain heart infusion (BHI) and plated onto five different agar media. All culture medias, except for MacConkey, were incubated anaerobically. Bacterial colonies were harvested in bulk and used for DNA extraction, 16S PCR amplification, and sequenced using the Illumina MiSeq platform. Predominant phyla in healthy and diseased horses were Firmicutes, followed by Bacteroidetes in all cultured medias, except for MacConkey agar, in which Proteobacteria was the dominant phylum. Greater bacterial richness was identified in sequenced cecal contents as compared to cultured plates (P < 0.05). Culture-enriched molecular profiling combined with 16S rRNA gene sequencing offer an alternative method for the study of the gut microbiota of horses. For direct cecum content 16S gene amplification, the alpha diversity indices were lower in diarrheic horses compared to healthy horses (P < 0.05). A higher relative abundance of Fusobacteriota was found in 2/6 samples from diarrheic horses. The role of Fusobacteriota in equine colitis deserves investigation.

Effect of plasma transfusion on serum amyloid A concentration in healthy neonatal foals and foals with failure of transfer of passive immunity.

BACKGROUND: Anecdotal evidence suggests plasma transfusions increase serum amyloid A (SAA) concentrations in healthy neonatal foals making this marker of inflammation inappropriate for therapeutic decision making in such animals. HYPOTHESIS/OBJECTIVES: Administration of hyperimmune fresh frozen plasma (FFP) increases SAA concentration in healthy foals and in foals with failure of transfer of passive immunity (FTPI). ANIMALS: Eighty-six healthy foals. METHODS: Prospective cohort study. Foals <24 hours of age receiving plasma transfusion for treatment of FTPI (serum immunoglobulin G [IgG] concentrations <8 g/L; n = 17) or as a preventative measure for Rhodococcus equi infection (IgG >8 g/L; n = 33) were enrolled. A healthy nontransfused group of foals (IgG >8 g/L; n = 21) also was included. Serum amyloid A concentration was determined before (t0h) and after (t24h) administration of FFP. Changes in blood SAA concentration were assessed using linear regression models. RESULTS: No statistical differences were found in SAA concentration at t0h or t24h among the 3 groups (P > .05, for all comparisons). The variation in SAA concentration before (t0h) and after (t24h) plasma transfusion showed that administration of FFP was not associated with the changes in SAA concentration (P > .05). An association between SAA concentration at t0h and at 24 hours (P < .05) was identified, where foals with higher SAA concentration at t0h also had higher SAA concentration at t24h. CONCLUSIONS AND CLINICAL IMPORTANCE: Administration of FFP to newborn foals was not associated with changes in SAA concentration.

The fecal bacterial microbiota of healthy and sick newborn foals.

BACKGROUND: The fecal bacterial microbiota of normal foals and foals with enterocolitis has been characterized using next-generation sequencing technology; however, there are no reports investigating the gut microbiota in foals hospitalized for other perinatal diseases. OBJECTIVE: To describe and compare the fecal bacterial microbiota in healthy and sick foals using next-generation sequencing techniques. ANIMALS: Hospitalized (17) and healthy foals (21). METHODS: Case-control study. Fecal samples were collected from healthy and sick foals on admission. Sick foals were further divided into sick nonseptic (SNS, n = 9) and septic (n = 8) foals. After extraction of DNA, the V4 region of the 16 S rRNA gene was amplified using a PCR assay, and the final product was sequenced with an Illumina MiSeq. RESULTS: Diversity was significantly lower in healthy than sick foals (P < .05). The bacterial membership (Jaccard index) and structure (Yue & Clayton index) of the fecal microbiota of healthy, septic, and SNS foals were similar (AMOVA, P > .05). Bacterial membership (AMOVA, P = .06) and structure (AMOVA, P = .33) were not different between healthy and sick foals. Enterobacteriaceae, Enterococcus, and Streptococcus were among the 5 more abundant taxa identified in both groups. CONCLUSION AND CLINICAL IMPORTANCE: Higher fecal microbiota diversity in sick than healthy foals might suggest a high exposure to environmental microorganisms or an unstable colonic microbiota. The presence of microorganisms causing bacteremia in foals in a high relative abundance in the feces of foals suggests the intestine might play an essential role in the causation of bacteremia in foals.

Fecal prevalence of Clostridium innocuum DNA in healthy horses and horses with colitis.

This study compared the prevalence of C. innocuum DNA in the feces of healthy horses and horses with acute colitis. C. innocuum was identified in 22% (15/68) of colitis cases and 18% (12/68) of healthy horses (p = 0.416).

Investigating fecal microbial transplant as a novel therapy in dogs with inflammatory bowel disease: A preliminary study.

BACKGROUND: There are limited studies investigating the use of fecal microbial transplant (FMT) in dogs with inflammatory bowel disease (IBD). The aim of this preliminary study was to assess the feasibility of adding FMT to standard therapy (corticosteroids and a hypoallergenic diet) for dogs with IBD and to and to describe the changes in measured outcomes after 30 days of treatment. METHODS: Thirteen client-owned dogs with IBD were enrolled in this double blinded, randomized clinical trial. All dogs received corticosteroid therapy and a hypoallergenic diet; dogs were randomized to receive either placebo or FMT. Measured outcomes included the canine chronic enteropathy clinical activity index (CCECAI) at 1 week and 1 month after enrolment. Fecal microbiota were analyzed after extracting DNA from fecal samples and profiling using 16S amplicon sequencing. Dogs in the placebo group not responding to treatment after 1 month were offered FMT. RESULTS: The CCECAI significantly decreased over time in both groups (p = 0.001). There were no significant differences between the CCECAI of the placebo and FMT group at each time point (F test from ANOVA, p = 0.40). No adverse effects were reported in the 30 days following FMT. CONCLUSIONS: The addition of FMT to standard therapy for IBD was feasible. No significant differences were observed in the CCECAI between groups at each time point. Large scale clinical trials can be performed using these methods to evaluate the longer term effect of FMT on clinical signs, microbial diversity, and other outcomes.

Fecal microbiota of horses with colitis and its association with laminitis and survival during hospitalization.

BACKGROUND: The association of microbiota with clinical outcomes and the taxa associated with colitis in horses remains generally unknown. OBJECTIVES: Describe the fecal microbiota of horses with colitis and investigate the association of the fecal microbiota with the development of laminitis and survival. ANIMALS: Thirty-six healthy and 55 colitis horses subdivided into laminitis (n = 15) and non-laminitis (n = 39, 1 horse with chronic laminitis was removed from this comparison) and survivors (n = 27) and nonsurvivors (n = 28). METHODS: Unmatched case-control study. The Illumina MiSeq platform targeting the V4 region of the 16S ribosomal RNA gene was used to assess the microbiota. RESULTS: The community membership (Jaccard index) and structure (Yue and Clayton index) were different (analysis of molecular variance [AMOVA]; P < .001) between healthy and colitis horses. The linear discriminant analysis effect size (LEfSe; linear discriminant analysis [LDA] >3; P < .05) and random forest analyses found Enterobacteriaceae, Lactobacillus, Streptococcus, and Enterococcus enriched in colitis horses, whereas Treponema, Faecalibacterium, Ruminococcaceae, and Lachnospiraceae were enriched in healthy horses. The community membership and structure of colitis horses with or without laminitis was (AMOVA; P > .05). Enterobacteriaceae, Streptococcus, and Lactobacillus were enriched in horses with laminitis (LDA > 3; P < .05). The community membership (AMOVA; P = .008) of surviving and nonsurviving horses was different. Nonsurviving horses had an enrichment of Enterobacteriaceae, Pseudomonas, Streptococcus, and Enterococcus (LDA >3; P < .05). CONCLUSION AND CLINICAL IMPORTANCE: Differences in the microbiota of horses with colitis that survive or do not survive are minor and, similarly, the microbiota differences in horses with colitis that do or do not develop laminitis are minor.