RESUMEN
Cryptosporidiosis has previously been reported in animals, humans, and water sources in the United Arab Emirates (UAE). However, most reports were only to the genus level, or generically identified as cryptosporidiosis. We aimed to investigate the genetic diversity of Cryptosporidium species occurring in diarrhetic ungulates which were brought to the Central Veterinary Research Laboratory (CVRL) in Dubai. Using a combination of microscopic and molecular methods, we identified five species of Cryptosporidium occurring among ungulates in the UAE, namely C. parvum, C. hominis, C. xiaoi, C. meleagridis, and C. equi. Cryptosporidium parvum was the most prevalent species in our samples. Furthermore, we identified subtypes of C. parvum and C. hominis, which are involved in both human and animal cryptosporidiosis. This is also the first reported occurrence of Cryptosporidium spp. in the Arabian Tahr, to our knowledge. Since the animals examined were all in contact with humans, the possibility of zoonotic spread is possible. Our study correlates with previous reports in the region, building upon the identification of Cryptosporidium sp. However, there is a need to further investigate the endemic populations of Cryptosporidium, including more hosts, sampling asymptomatic animals, and location data.
Asunto(s)
Criptosporidiosis , Cryptosporidium , Diarrea , Variación Genética , Emiratos Árabes Unidos/epidemiología , Animales , Criptosporidiosis/parasitología , Criptosporidiosis/epidemiología , Cryptosporidium/genética , Cryptosporidium/clasificación , Cryptosporidium/aislamiento & purificación , Diarrea/veterinaria , Diarrea/parasitología , Diarrea/epidemiología , Heces/parasitología , Bovinos , Filogenia , Cabras/parasitología , ADN Protozoario/genéticaRESUMEN
Porcine epidemic diarrhea virus (PEDV) causes diarrhea and vomiting in piglets, leading to a mortality rate of 100%. Due to the high frequency of mutation, it is important to monitor the evolution of PEDV and develop potential vaccine candidates. In this study, two PEDV strains (ZJ2022 and ZQ2022) were identified by PCR. These strains were subsequently isolated, and their genome sequences, growth characteristics, and pathogenicity were compared. Phylogenetic and recombination analyses revealed that both strains belonged to GIIa-subgroup, and ZQ2022 was identified as a recombinant strain derived from ZJ2022. Further sequence analysis showed that the ZJ2022 strain had a modified top region of the S1 protein due to a three amino acid insertion (T380_Y380insGGE) in the S1 gene. According to the virus growth curve, ZJ2022 exhibited better cellular adaptation than ZQ2022, with higher viral titers from 8 hpi to 24 hpi. Additionally, ZQ2022 exhibited a high level of pathogenicity, causing severe diarrhea in piglets at 36 hpi and a 100% mortality rate by 96 hpi. In contrast, ZJ2022 showed lower pathogenicity, inducing severe diarrhea in piglets at 60 hpi, with a mortality rate of 60% at 96 hpi and 100% at 120 hpi. In summary, our findings provided evidence of the undergoing mutations in Chinese PEDV strains. Furthermore, the S gene insertion strain ZJ2022 exhibited strong cellular adaptability and low pathogenicity, making it a potential candidate strain for vaccine development.
Asunto(s)
Animales Recién Nacidos , Infecciones por Coronavirus , Diarrea , Filogenia , Virus de la Diarrea Epidémica Porcina , Enfermedades de los Porcinos , Animales , Virus de la Diarrea Epidémica Porcina/genética , Virus de la Diarrea Epidémica Porcina/patogenicidad , Virus de la Diarrea Epidémica Porcina/aislamiento & purificación , Virus de la Diarrea Epidémica Porcina/clasificación , Porcinos , Enfermedades de los Porcinos/virología , Infecciones por Coronavirus/veterinaria , Infecciones por Coronavirus/virología , Virulencia , Diarrea/virología , Diarrea/veterinaria , Glicoproteína de la Espiga del Coronavirus/genética , Genoma Viral , Mutagénesis Insercional , China , Células VeroRESUMEN
BACKGROUND: Calf diarrhea is a major cause of morbidity and mortality in the livestock sector worldwide and it can be caused by multiple infectious agents. In Ethiopia, cattle are the most economically important species within the livestock sector, but at the same time the young animals suffer from high rates of morbidity and mortality due to calf diarrhea. However, studies including both screening and molecular characterization of bovine enteric pathogens are lacking. Therefore, we aimed to both detect and molecularly characterize four of the major enteric pathogens in calf diarrhea, Enterotoxigenic Escherichia coli (E. coli K99 +), Cryptosporidium spp., rotavirus A (RVA), and bovine coronavirus (BCoV) in calves from central Ethiopia. Diarrheic and non-diarrheic calves were included in the study and fecal samples were analyzed with antigen-ELISA and quantitative real-time PCR (qPCR). Positive samples were further characterized by genotyping PCRs. RESULTS: All four pathogens were detected in both diarrheic and non-diarrheic calves using qPCR and further characterization showed the presence of three Cryptosporidium species, C. andersoni, C. bovis and C. ryanae. Furthermore, genotyping of RVA-positive samples found a common bovine genotype G10P[11], as well as a more unusual G-type, G24. To our knowledge this is the first detection of the G24 RVA genotype in Ethiopia as well as in Africa. Lastly, investigation of the spike gene revealed two distinct BCoV strains, one classical BCoV strain and one bovine-like CoV strain. CONCLUSIONS: Our results show that Cryptosporidium spp., E. coli K99 + , RVA and BCoV circulate in calves from central Ethiopia. Furthermore, our findings of the rare RVA G-type G24 and a bovine-like CoV demonstrates the importance of genetic characterization.
Asunto(s)
Enfermedades de los Bovinos , Coronavirus Bovino , Cryptosporidium , Diarrea , Heces , Rotavirus , Animales , Bovinos , Etiopía/epidemiología , Diarrea/veterinaria , Diarrea/virología , Diarrea/microbiología , Diarrea/parasitología , Enfermedades de los Bovinos/virología , Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/microbiología , Enfermedades de los Bovinos/parasitología , Heces/virología , Heces/parasitología , Heces/microbiología , Rotavirus/genética , Rotavirus/aislamiento & purificación , Rotavirus/clasificación , Cryptosporidium/aislamiento & purificación , Cryptosporidium/genética , Cryptosporidium/clasificación , Coronavirus Bovino/genética , Coronavirus Bovino/aislamiento & purificación , Escherichia coli Enterotoxigénica/aislamiento & purificación , Escherichia coli Enterotoxigénica/genética , Genotipo , Criptosporidiosis/epidemiología , Infecciones por Rotavirus/veterinaria , Infecciones por Rotavirus/epidemiología , Infecciones por Rotavirus/virología , Infecciones por Coronavirus/veterinaria , Infecciones por Coronavirus/virología , Infecciones por Coronavirus/epidemiología , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Infecciones por Escherichia coli/veterinaria , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/microbiologíaRESUMEN
Dihydromyricetin (DMY), a natural flavonoid compound, are believed to prevent inflammatory response, dealing with pathogens and repairing the intestinal barrier. The objective of this study was to investigate whether DMY supplementation could attenuate intestinal damage in the context of enterotoxigenic Escherichia coli K88 (ETEC F4+) infection. After weaning, different litters of pigs were randomly assigned to one of the following treatments: (1) non-challenged control (CON, fed with basal diet); (2) ETEC-challenged control (ECON, fed with basal diet); and (3) ETEC challenge + DMY treatment (EDMY, fed with basal diet plus 300 mg kg-1 DMY). We observed a significant reduction in fecal Escherichia coli shedding and diarrhea incidence, but an increase in ADG in pigs of EDMY group compared to the pigs of ECON group. Relative to the pigs of ECON group, dietary DMY treatment decreased (P < 0.05) concentrations of the serum D-xylose, D-lactate and diamine oxidase (DAO), but increased the abundance of zonula occludens-1 (ZO-1) in the jejunum of pigs. In addition, DMY also decreased (P < 0.05) the number of S-phase cells and the percentage of total apoptotic epithelial cells of jejunal epithelium in pigs of the EDMY group compared to the pigs of the ECON group. Furthermore, DMY decreased the mRNA expression levels of critical immune-associated genes TLR4, NFκB, Caspase3, Caspase9, IL-1ß, IL-6, TNF-α and the protein p-NFκB and p-IκBα expressions of intestinal epithelium in pigs of the EDMY group compared to the pigs of the ECON group. Compared to the ECON group, DMY elevated (P < 0.05) the expression levels of ß-defensins PBD1, PBD2, PBD3, PBD129, as well as the abundance of secreted IgA in intestinal mucosae of the EDMY group. Thus, our results indicate that DMY may relieve intestinal integrity damage due to Escherichia coli F4.
Asunto(s)
Escherichia coli Enterotoxigénica , Infecciones por Escherichia coli , Flavonoles , Mucosa Intestinal , Enfermedades de los Porcinos , Animales , Escherichia coli Enterotoxigénica/efectos de los fármacos , Porcinos , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/patología , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiología , Mucosa Intestinal/inmunología , Flavonoles/farmacología , Flavonoles/uso terapéutico , Infecciones por Escherichia coli/tratamiento farmacológico , Infecciones por Escherichia coli/veterinaria , Infecciones por Escherichia coli/inmunología , Enfermedades de los Porcinos/tratamiento farmacológico , Enfermedades de los Porcinos/microbiología , Enfermedades de los Porcinos/inmunología , Destete , Citocinas/metabolismo , Diarrea/tratamiento farmacológico , Diarrea/veterinaria , Apoptosis/efectos de los fármacos , Proteína de la Zonula Occludens-1/metabolismo , Proteína de la Zonula Occludens-1/genéticaRESUMEN
Snakes are sometimes regarded as pets and are used in traditional Chinese medicine. Cryptosporidium spp. are frequently identified in snakes, representing an important pathogen and causing gastrointestinal diseases. Current data indicate that risk factors for infection and patterns of clinical symptom presentation may differ among Cryptosporidium spp. To better understand the infection status by Cryptosporidium spp., fecal samples were collected from 603 asymptomatic and 147 symptomatic snakes in 26 provinces of China. These samples came from Elaphe guttata, Elaphe obsoleta, Pituophis melanoleucus, Thamnophis sirtalis, Lampropeltis getulus, and Heterodon nasicus. The partial small subunit (SSU) rRNA gene was amplified using nested polymerase chain reaction (PCR) to investigate the infection rate of Cryptosporidium spp., and to assess evolutionary relationships and genetic characterization. A prevalence of 20% was recorded in asymptomatic snakes, with age identified as a significant risk factor. In contrast, 70% of symptomatic snakes were positive for Cryptosporidium spp., with Cryptosporidium serpentis and Cryptosporidium varanii (syn. C. saurophilum). Further analysis revealed a potential association between C. serpentis and regurgitation, and C. varanii and diarrhea, while neither species was linked to flatulence. To our knowledge, this is the first study to report Cryptosporidium spp. and associated clinical signs in symptomatic snakes in China. This study aims to enhance the understanding of Cryptosporidium infections, risk factors, and clinical manifestations in snakes, providing data crucial for the control and prevention of cryptosporidiosis.
Title: Cryptosporidium spp. chez les serpents captifs de 26 provinces de Chine : prévalence, caractérisation moléculaire et symptômes. Abstract: Les serpents sont parfois considérés comme animaux de compagnie et sont utilisés en médecine traditionnelle chinoise. Des Cryptosporidium spp. sont fréquemment identifiés chez les serpents, ont un rôle d'agent pathogène important et provoquent des maladies gastro-intestinales. Les données actuelles indiquent que les facteurs de risque d'infection et les schémas de présentation des symptômes cliniques peuvent varier en fonction des espèces de Cryptosporidium. Pour mieux comprendre l'état d'infection par Cryptosporidium spp., des échantillons fécaux ont été collectés auprès de 603 serpents asymptomatiques et 147 serpents symptomatiques dans 26 provinces de Chine. Ces échantillons provenaient d'Elaphe guttata, Elaphe obsoleta, Pituophis melanoleucus, Thamnophis sirtalis, Lampropeltis getulus et Heterodon nasicus. Le gène de l'ARNr de la petite sous-unité partielle (SSU) a été amplifié à l'aide d'une réaction en chaîne par polymérase (PCR) imbriquée pour étudier le taux d'infection par Cryptosporidium spp. et évaluer les relations évolutives et la caractérisation génétique. Une prévalence de 20 % a été trouvée chez les serpents asymptomatiques, l'âge étant identifié comme un facteur de risque important. En revanche, 70 % des serpents symptomatiques étaient positifs à Cryptosporidium spp. avec Cryptosporidium serpentis et Cryptosporidium varanii (syn. C. saurophilum). Une analyse plus approfondie a révélé une association potentielle entre C. serpentis et la régurgitation, et C. varanii et la diarrhée, alors qu'aucune des deux espèces n'était liée aux flatulences. À notre connaissance, il s'agit ici de la première étude à signaler la présence de Cryptosporidium spp. et les signes cliniques associés chez des serpents symptomatiques en Chine. Cette étude vise à améliorer la compréhension des infections à Cryptosporidium, des facteurs de risque et des manifestations cliniques chez les serpents, en fournissant des données cruciales pour le contrôle et la prévention de la cryptosporidiose.
Asunto(s)
Criptosporidiosis , Cryptosporidium , Heces , Serpientes , Animales , Cryptosporidium/genética , Cryptosporidium/aislamiento & purificación , Cryptosporidium/clasificación , Criptosporidiosis/epidemiología , Criptosporidiosis/parasitología , China/epidemiología , Prevalencia , Heces/parasitología , Serpientes/parasitología , Filogenia , Factores de Riesgo , Reacción en Cadena de la Polimerasa/veterinaria , Masculino , Femenino , ADN Protozoario/aislamiento & purificación , Diarrea/parasitología , Diarrea/veterinaria , Diarrea/epidemiología , Mascotas/parasitologíaRESUMEN
The objective of this study was to examine the values of MX dynamin-like GTPase 1 (Mx1), high mobility group box-1 (HMGB1), systemic inflammatory response index (SIRI), systemic inflammatory index (SII), tumor necrosis factor (TNF), and other hematological indices in calves with systemic inflammatory response syndrome (SIRS). The study material was divided into two groups: the SIRS group (comprising 13 calves) and the control group (comprising 10 calves). The independent samples t-test and Mann-Whitney U test were employed for normally distributed and non-normally distributed data, respectively. The relationship between the two groups was determined using Spearman correlation coefficient analysis. Significant differences were identified between the SIRS group and the control group with regard to white blood cell (WBC; P < 0.05), neutrophil (NEU; P < 0.01), and neutrophil-to-lymphocyte ratio (NLR; P < 0.001) values, in addition to SIRI (P < 0.05), SII (P < 0.01) values. Furthermore, HMGB1 (P < 0.001), Mx1 (P < 0.05), and TNF values (P < 0.001) demonstrated notable disparities between the two groups. As a result of this study, it was concluded that there were significant increases in inflammatory hematological indices, as well as in the levels of HMGB1, Mx1, and TNF, in calves with SIRS.
Asunto(s)
Animales Recién Nacidos , Enfermedades de los Bovinos , Diarrea , Proteína HMGB1 , Proteínas de Resistencia a Mixovirus , Síndrome de Respuesta Inflamatoria Sistémica , Factor de Necrosis Tumoral alfa , Animales , Proteína HMGB1/sangre , Bovinos , Síndrome de Respuesta Inflamatoria Sistémica/veterinaria , Síndrome de Respuesta Inflamatoria Sistémica/sangre , Síndrome de Respuesta Inflamatoria Sistémica/inmunología , Proteínas de Resistencia a Mixovirus/genética , Factor de Necrosis Tumoral alfa/sangre , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/sangre , Animales Recién Nacidos/inmunología , Diarrea/veterinaria , Diarrea/inmunología , Masculino , Femenino , Inflamación/veterinaria , Inflamación/sangre , Inflamación/inmunologíaRESUMEN
BACKGROUND: Cupriavidus gilardii is an aerobic, gram-negative, motile, glucose-nonfermenting bacillus, first described in 1999. Typically, it exhibits low pathogenicity in humans, causing opportunistic infections primarily in individuals with compromised immune systems. This bacterium has been also found in various environmental sources such as plants and contaminated soils. Notably, there have been no documented cases of C. gilardii infections in animals. CASE PRESENTATION: This case report outlines a bovine neonatal diarrhea outbreak that occurred in Northern Greece, during which C. gilardii was isolated. Faecal samples from 5-day-old calves were collected and transported to the laboratory for further examination. Bacterial culture and next generation sequencing techniques were employed to confirm the presence of this bacterium in the samples. Following the isolation and identification of C. gilardii from the samples, an autogenous vaccine was produced and administered to the cows within the farm. Subsequent to vaccination, a progressive reduction in calf diarrhea and deaths was observed, leading to their eventual complete resolution. To the best of our knowledge, this represents the first documentation of C. gilardii isolation from cases of bovine neonatal diarrhea. CONCLUSION: This case report presents the first isolation case of C. gilardii from animal samples and more specifically from calf faecal samples. It represents an important observation, providing evidence that this opportunistic human pathogen could contribute to clinical symptoms in animals.
Asunto(s)
Animales Recién Nacidos , Enfermedades de los Bovinos , Cupriavidus , Diarrea , Brotes de Enfermedades , Heces , Infecciones por Bacterias Gramnegativas , Animales , Bovinos , Diarrea/veterinaria , Diarrea/microbiología , Diarrea/epidemiología , Brotes de Enfermedades/veterinaria , Enfermedades de los Bovinos/microbiología , Enfermedades de los Bovinos/epidemiología , Heces/microbiología , Infecciones por Bacterias Gramnegativas/veterinaria , Infecciones por Bacterias Gramnegativas/epidemiología , Infecciones por Bacterias Gramnegativas/microbiología , Cupriavidus/aislamiento & purificación , Cupriavidus/genética , Grecia/epidemiologíaRESUMEN
Porcine epidemic diarrhea virus (PEDV) is an enteric coronavirus that has been the main cause of diarrhea in piglets since 2010 in China. The aim of this study was to investigate sequence variation and recombination events in the spike (S) gene of PEDV isolates from China. Thirty complete S gene sequences were obtained from PEDV-positive samples collected in six provinces in China from 2020 to 2023. Phylogenetic analysis showed that 10% (3/30) belonged to subtype GII-a, 6.67% (2/30) were categorized as subtype GII-b, 66.67% (20/30) were categorized as subtype GII-c, and 16.66% (5/30) were clustered with the S-INDEL strains. Amino acid sequence alignments showed that, when compared to strains of other subtypes, the GII-c strains had two characteristic amino acid substitutions (N139D and I289M). Five S-INDEL subtype strains had a single amino acid deletion (139N) and four amino acid substitutions (N118G, T137S, A138S, and D141G). Recombination analysis allowed six putative recombination events to be identified, one involving recombination between GII-c strains, two involving GII-c and GII-b strains, two involving GII-c and GI-a strains, and one involving GII-a and GI-b strains. These results suggest that recombination between PEDV strains has been common and complex in recent years and is one of the main reasons for the continuous variation of PEDV strains.
Asunto(s)
Infecciones por Coronavirus , Virus de la Diarrea Epidémica Porcina , Recombinación Genética , Glicoproteína de la Espiga del Coronavirus , Enfermedades de los Porcinos , Animales , Secuencia de Aminoácidos , Sustitución de Aminoácidos , China/epidemiología , Infecciones por Coronavirus/virología , Infecciones por Coronavirus/veterinaria , Infecciones por Coronavirus/epidemiología , Diarrea/virología , Diarrea/veterinaria , Diarrea/epidemiología , Variación Genética , Genotipo , Filogenia , Virus de la Diarrea Epidémica Porcina/genética , Virus de la Diarrea Epidémica Porcina/clasificación , Virus de la Diarrea Epidémica Porcina/aislamiento & purificación , Glicoproteína de la Espiga del Coronavirus/genética , Porcinos , Enfermedades de los Porcinos/virología , Enfermedades de los Porcinos/epidemiologíaRESUMEN
Porcine epidemic diarrhea virus (PEDV), porcine deltacoronavirus (PDCoV) and transmissible gastroenteritis virus (TGEV) are three clinically common coronaviruses causing diarrhea in pigs, with indistinguishable clinical signs and pathological changes. Rapid, portable and reliable differential diagnosis of these three pathogens is crucial for the prompt implementation of appropriate control measures. In this study, we developed a triplex nucleic acid assay that combines reverse transcription recombinase-aided amplification (RT-RAA) with lateral flow assay (LFA) by targeting the most conserved genomic region in the ORF1b genes of PEDV, PDCoV and TGEV. The entire detection process of the triplex RT-RAA-LFA assay included 10-min nucleic acid amplification at 42 °C and 5-min visual LFA readout at room temperature. The assay could specifically differentiate PEDV, PDCoV and TGEV without cross-reaction with any other major swine pathogens. Sensitivity analysis showed that the triplex RT-RAA-LFA assay was able to detect the viral RNA extracted from the spiked fecal samples with the minimum of 1 × 100 TCID50 PEDV, 1 × 104 TCID50 PDCoV, and 1 × 102 TCID50 TGEV per reaction, respectively. Further analysis showed that the 95 % detection limit (LOD) of triplex RT-RAA-LFA for PEDV, PDCoV, and TGEV were 22, 478, and 205 copies of recombinant plasmids per reaction, respectively. The diagnostic performance of triplex RT-RAA-LFA was compared with that of PEDV, PDCoV and TGEV respective commercial real-time RT-PCR kits by testing 114 clinical rectal swab samples in parallel. The total diagnostic coincidence rates of triplex RT-RAA-LFA with real-time RT-PCR kits of PEDV, PDCoV and TGEV were 100 %, 99.1 % and 99.1 %, respectively, and their Kappa values were 1.00, 0.958 and 0.936, respectively. Collectively, the RT-RAA-LFA assay is a powerful tool for the rapid, portable, visual, and synchronous differential diagnosis of PEDV, PDCoV, and TGEV.
Asunto(s)
Infecciones por Coronavirus , Deltacoronavirus , Heces , Técnicas de Amplificación de Ácido Nucleico , Virus de la Diarrea Epidémica Porcina , ARN Viral , Sensibilidad y Especificidad , Enfermedades de los Porcinos , Virus de la Gastroenteritis Transmisible , Animales , Porcinos , Virus de la Gastroenteritis Transmisible/aislamiento & purificación , Virus de la Gastroenteritis Transmisible/genética , Virus de la Diarrea Epidémica Porcina/aislamiento & purificación , Virus de la Diarrea Epidémica Porcina/genética , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/virología , ARN Viral/genética , ARN Viral/aislamiento & purificación , Heces/virología , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/veterinaria , Infecciones por Coronavirus/virología , Diagnóstico Diferencial , Deltacoronavirus/aislamiento & purificación , Deltacoronavirus/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Técnicas de Amplificación de Ácido Nucleico/veterinaria , Gastroenteritis Porcina Transmisible/diagnóstico , Gastroenteritis Porcina Transmisible/virología , Técnicas de Diagnóstico Molecular/métodos , Diarrea/virología , Diarrea/veterinaria , Diarrea/diagnósticoRESUMEN
A total of 490 diarrhoeic samples from calves aged between 0 and 6 months were screened for the presence of different G- and P-genotypes of rotavirus circulating in bovines in the Kashmir Valley. Of the 490 diarrhoeic samples, Group A rotavirus was detected in 68 (13.87%) samples by polymerase chain reaction (PCR) followed by RNA-PAGE. Genotyping analysis revealed G10, G6, G3, P[11] and P[5] to be the predominant types. The most common types of combinations detected were G10P[11] (27.90%) and G6P[11] (20.60%). The prevalence rate of G10 and P[11] decreased from 60% to 36.76% and 100%-69.11%, respectively. Genotypes G6, G3, P[1] and P[5], which were not previously reported, were detected and unusual combinations such as G6P[11], G3P[11], G10P[5], G3P[5], G6P[1], G6P[5], G6+G8P[11] were also observed for the first time. Fluctuations in the predominant types, emergence of new types and possible genetic reassortment events suggest an unstable epidemiological situation and the need for continuous surveillance of the circulating types to ensure the suitability of the vaccination programme. The present data suggests G10, G6, P[11] and P[5] genotypes could be incorporated in the polyvalent vaccine to offer increased protection against bovine rotavirus infection in India.
Asunto(s)
Enfermedades de los Bovinos , Diarrea , Genotipo , Infecciones por Rotavirus , Rotavirus , Animales , Rotavirus/genética , Rotavirus/clasificación , Bovinos , India/epidemiología , Infecciones por Rotavirus/veterinaria , Infecciones por Rotavirus/epidemiología , Infecciones por Rotavirus/virología , Enfermedades de los Bovinos/virología , Enfermedades de los Bovinos/epidemiología , Diarrea/veterinaria , Diarrea/virología , Diarrea/epidemiología , Animales Recién NacidosRESUMEN
This study investigates the presence of virulence-associated genes and antimicrobial resistance (AMR) in Escherichia fergusonii isolates obtained from ostrich chicks. A total of 287 isolates were recovered from 106 fecal samples from ostrich chicks suffering from diarrhea and subjected to molecular identification and biochemical characterization. E. fergusonii was detected in 10 samples (9.4â¯%) using two PCR-detection protocols. Notably, the isolates lacked various virulence genes commonly associated with pathogenic E. coli including elt, est, stx, eae, ehly, cdt, iss, iutA, iroN, hlyA, ompT, except for one isolate harboring the astA gene. Antimicrobial susceptibility testing revealed that all isolates were susceptible to ciprofloxacin, while high resistance was observed against amoxicillin clavulanate (AMC), trimethoprim-sulfamethoxazole (SXT), and doxycycline (D). Moreover, eight isolates displayed multidrug resistance (MDR) and four exhibited resistance to 9-11 antimicrobials. The most frequent resistance gene was sul2, which was present in all isolates; the other resistance genes detected consisted of int1 (4/10), int2 (3/10), blaCMY (2/10), and qnrS, blaTEM, blaCMY, blaCTX-M, and flo each were detected only in one E. fergusonii Isolate. Plasmid replicon typing identified the presence of I1 (7/10), N (5/10), and Y (1/10). This study provides valuable insights into the virulence and antimicrobial resistance of E. fergusonii isolates from ostrich chicks, highlighting the complexity of antimicrobial resistance mechanisms exhibited by these bacteria. Further research is essential to understand the transmission dynamics and clinical implications of these findings in veterinary and public health settings.
Asunto(s)
Antibacterianos , Escherichia , Heces , Pruebas de Sensibilidad Microbiana , Struthioniformes , Animales , Struthioniformes/microbiología , Antibacterianos/farmacología , Escherichia/genética , Escherichia/efectos de los fármacos , Escherichia/patogenicidad , Escherichia/aislamiento & purificación , Heces/microbiología , Virulencia/genética , Factores de Virulencia/genética , Farmacorresistencia Bacteriana Múltiple/genética , Diarrea/microbiología , Diarrea/veterinaria , Infecciones por Enterobacteriaceae/veterinaria , Infecciones por Enterobacteriaceae/microbiología , Farmacorresistencia Bacteriana/genética , Enfermedades de las Aves/microbiología , Genes Bacterianos/genéticaRESUMEN
Cryptosporidiosis is a parasitic disease caused by the protozoan Cryptosporidium in vertebrates. In livestock, especially ruminants, infants develop diarrheal syndromes. The infection is common worldwide , including Iran, where it is reported in several species. Morphological diagnosis of Cryptosporidium species is associated with many limitations and has no taxonomic value on its own, so the use of molecular methods can overcome these limitations to some extent. The present aims at microscopic, molecular and antigen detection and isolation of Cryptosporidium parvum parasites. Firstly, 300 samples were collected from different parts of Iran. Subsequently oocysts from feces were purified by the method of Casemore et al. using the flotation technique and stained by the modified Ziehl-Neelsen method (Henriksen method) and identified by diagnostic keys. ELISA test was also performed on the samples with results ranging from 1 to 4 positive. The results of our study show that, of the 300 cases tested for Cryptosporidium, 48 cases (16%) and 54 (18%) were positive in ELISA and PCR, respectively. Microscopic evaluation also mainly confirmed the ELISA results. These cases were collected in summer, autumn, and winter, with, more than 50% of the positive cases found among the samples collected in autumn. In addition, 54 positive cases were found by PCR test, which is 6 cases more than ELISA results. Finally, the results of PCR detection and ELISA were subjected to chi-square analysis, where no significant difference was found between the collected data (p=0.0587).
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Enfermedades de los Bovinos , Criptosporidiosis , Cryptosporidium parvum , Diarrea , Ensayo de Inmunoadsorción Enzimática , Criptosporidiosis/parasitología , Criptosporidiosis/epidemiología , Cryptosporidium parvum/aislamiento & purificación , Irán/epidemiología , Animales , Enfermedades de los Bovinos/parasitología , Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/diagnóstico , Bovinos , Ensayo de Inmunoadsorción Enzimática/veterinaria , Diarrea/veterinaria , Diarrea/parasitología , Diarrea/epidemiología , Antígenos de Protozoos/análisis , Reacción en Cadena de la Polimerasa/veterinaria , Heces/parasitologíaRESUMEN
Equine rotavirus species A (ERVA) G3P[12] and G14P[12] are two dominant genotypes that cause foal diarrhoea with a significant economic impact on the global equine industry. ERVA can also serve as a source of novel (equine-like) rotavirus species A (RVA) reassortants with zoonotic potential as those identified previously in 2013-2019 when equine G3-like RVA was responsible for worldwide outbreaks of severe gastroenteritis and hospitalizations in children. One hurdle to ERVA research is that the standard cell culture system optimized for human rotavirus replication is not efficient for isolating ERVA. Here, using an engineered cell line defective in antiviral innate immunity, we showed that both equine G3P[12] and G14P[12] strains can be rapidly isolated from diarrhoeic foals. The genome sequence analysis revealed that both G3P[12] and G14P[12] strains share the identical genotypic constellation except for VP7 and VP6 segments in which G3P[12] possessed VP7 of genotype G3 and VP6 of genotype I6 and G14P[12] had the combination of VP7 of genotype G14 and VP6 of genotype I2. Further characterization demonstrated that two ERVA genotypes have a limited cross-neutralization. The lack of an in vitro broad cross-protection between both genotypes supported the increased recent diarrhoea outbreaks due to equine G14P[12] in foals born to dams immunized with the inactivated monovalent equine G3P[12] vaccine. Finally, using the structural modelling approach, we provided the genetic basis of the antigenic divergence between ERVA G3P[12] and G14P[12] strains. The results of this study will provide a framework for further investigation of infection biology, pathogenesis and cross-protection of equine rotaviruses.
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Antígenos Virales , Diarrea , Genotipo , Enfermedades de los Caballos , Infecciones por Rotavirus , Rotavirus , Animales , Caballos , Rotavirus/genética , Rotavirus/inmunología , Rotavirus/aislamiento & purificación , Rotavirus/clasificación , Infecciones por Rotavirus/veterinaria , Infecciones por Rotavirus/virología , Infecciones por Rotavirus/inmunología , Enfermedades de los Caballos/virología , Enfermedades de los Caballos/inmunología , Diarrea/virología , Diarrea/veterinaria , Antígenos Virales/genética , Antígenos Virales/inmunología , Genoma Viral/genética , Filogenia , Línea CelularRESUMEN
BACKGROUND: Calf diarrhea leads to high mortality rates and decreases in growth and productivity, causing negative effects on the livestock industry. Lactate is closely associated with metabolic acidosis in diarrheic calves. However, there have been no reports on lactate concentrations in Korean indigenous (Hanwoo) calves, especially those with diarrhea. This study aimed to determine the reference range of L-lactate and D-lactate concentrations in Hanwoo calves and to better understand the utility of lactate as predictive factors for acidemia in diarrheic calves. RESULTS: L-lactate and D-lactate concentrations were measured in healthy (n = 44) and diarrheic (n = 93) calves, and blood gas analysis was performed on diarrheic calves. The reference range in healthy calves was 0.2-2.25 mmol/L for L-lactate and 0.42-1.38 mmol/L for D-lactate. Diarrheic calves had higher concentrations of L-lactate and D-lactate than healthy calves. In diarrheic calves, L-lactate and D-lactate each had weak negative correlation with pH (r = - 0.31 and r = - 0.35). In diarrheic calves with hyper-L-lactatemia, the combined concentrations of L-lactate and D-lactate had moderate correlation with pH (r = - 0.51) and anion gap (r = 0.55). Receiver operating characteristic analysis showed D-lactate had fair predictive performance (AUC = 0.74) for severe acidemia, with an optimal cut-off value of > 1.43 mmol/L. The combined concentrations of L-lactate and D-lactate showed fair predictive performance for predicting acidemia (AUC = 0.74) and severe acidemia (AUC = 0.72), with cut-off values of > 6.05 mmol/L and > 5.95 mmol/L. CONCLUSIONS: The determined reference ranges for L-lactate and D-lactate in Hanwoo calves enable the identification of hyper-L-lactatemia and hyper-D-lactatemia. Diarrheic calves exhibited increased lactate concentrations correlated with acid-base parameters. While the concentrations of L-lactate and D-lactate have limitations as single diagnostic biomarkers for predicting acidemia or severe acidemia, their measurement remains important, and L-lactate has the advantage of being measurable at the point-of-care. Assessing lactate concentrations should be considered by clinicians, especially when used alongside other clinical indicators and diagnostic tests. This approach can improve calf diarrhea management, contributing positively to animal welfare and providing economic benefits to farms.
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Acidosis , Enfermedades de los Bovinos , Diarrea , Ácido Láctico , Animales , Bovinos , Diarrea/veterinaria , Diarrea/sangre , Enfermedades de los Bovinos/sangre , Enfermedades de los Bovinos/diagnóstico , Ácido Láctico/sangre , Acidosis/veterinaria , Acidosis/sangre , Acidosis/diagnóstico , República de Corea , Masculino , Femenino , Valores de Referencia , Análisis de los Gases de la Sangre/veterinariaRESUMEN
This study aimed to investigate the mechanism of quercetin increasing growth performance and decreasing incidence of diarrhea in weaned piglets. Forty-eight Duroc × Landrace × Large White weaned piglets with similar body weight (7.48 ± 0.20 kg, 28 days of age) were randomly divided into four treatments (control, 250 mg/kg quercetin, 500 mg/kg quercetin, and 750 mg/kg quercetin treatments) and fed with basal diet or experimental diet supplemented with quercetin. Performance, diarrhea rate and index, and content of serum anti-inflammatory factors were determined and calculated in weaned piglets; colonic flora and signaling pathways related to anti-inflammation were measured using 16S rDNA sequencing and RNA-seq, respectively. The results showed that compared with control, feed-to-gain ratio and content of serum interferon gamma (IFN-γ) were significantly decreased in the 500 and 750 mg/kg quercetin treatments (P < 0.05); quercetin significantly decreased diarrhea rate and diarrhea index (P < 0.05) and significantly increased the content of serum transforming growth factor (TGF-ß) in weaned piglets (P < 0.05); the content of serum NF-κB was significantly decreased in the 750 mg/kg quercetin treatment (P < 0.05); moreover, quercetin significantly increased diversity of colonic flora (P < 0.05), and at the phylum level, the relative abundance of Actinobacteria in the 500 and 750 mg/kg treatments was significantly increased (P < 0.05), and the relative abundance of Proteobacteria in the three quercetin treatments were significantly decreased (P < 0.05) in the colon of weaned piglets; at the genus level, the relative abundance of Clostridium-sensu-stricto-1, Turicibacter, unclassified_f_Lachnospiraceae, Phascolarctobacterium, and Family_XIII _AD3011_group was significantly increased (P < 0.05); the relative abundance of Subdollgranulum and Blautia was significantly decreased in the 500 and 750 mg/kg treatments (P < 0.05); the relative abundance of Eschericha-Shigella, Terrisporobacter, and Eubacterium-coprostanoligenes was significantly increased (P < 0.05); the relative abundance of Streptocococcus, Sarcina, Staphylococcus, and Ruminococcaceae_UCG-008 was significantly decreased in the three quercetin treatments (P < 0.05); the relative abundance of Ruminococcaceae_UCG_014 was significantly increased in the 250 mg/kg quercetin treatment in the colon of weaned piglets (P < 0.05). The results of Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that differentially expressed genes (DEGs) from the quercetin treatments were significantly enriched in nuclear transcription factor-κB (NF-κB) signal pathway (P < 0.05); mRNA expression of tumor necrosis factor-α (TNF-α), interleukin-1R1 (IL-1R1), conserved helix-loop-helix ubiquitous kinase (CHUK), toll-like receptor 4 (TLR4), and IL-1ß from quercetin treatments were significantly decreased in colonic mucosa of weaned piglets (P < 0.05). In summary, quercetin increased feed conversion ratio and decreased diarrhea through regulating NF-κB signaling pathway, controlling the balance between anti-inflammatory and proinflammatory factors, and modulating intestinal flora, thus promoting the absorption of nutrients in weaned piglets. These results provided the theoretical foundation for applying quercetin in preventing weaning piglets' diarrhea and animal husbandry practices.
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Diarrea , Quercetina , Destete , Animales , Quercetina/farmacología , Quercetina/uso terapéutico , Porcinos , Diarrea/veterinaria , Diarrea/tratamiento farmacológico , Microbioma Gastrointestinal/efectos de los fármacos , Enfermedades de los Porcinos/microbiología , Enfermedades de los Porcinos/tratamiento farmacológico , IncidenciaRESUMEN
Diarrhea is a common issue in domestic yaks (Bos grunniens) that can occur with pasture alterations and significantly impacts growth performance. Previous research has examined the microbiota of diarrhetic yaks; however, the structural changes in gut bacterial community and microbial interactions in yaks with grassland alteration-induced diarrhea remain poorly understood. To explore variations in gut microbiota homeostasis among yaks suffering from diarrhea, fecal microbiota diversity and composition were analyzed using 16 S rRNA amplicon sequencing. Gut fecal microbiota diversity was lower in diarrhetic yaks than in non-diarrhetic yaks. Furthermore, the bacterial community composition (including that of Proteobacteria and Actinobacteria) in the feces of diarrhetic yaks displayed significant alterations. Co-occurrence network analysis further underscored the compromised intestinal flora stability in yaks with diarrhea relative to that in non-diarrhetic yaks. Interestingly, the abundance of beneficial bacteria, such as Lachnospiraceae_AC2044_group and Lachnospiraceae_NK4A136_group, were decreased in yaks with diarrhea, and the reductions were negatively correlated with the fecal water content. Collectively, these findings indicate that diminished microbial stability and increased abundance of certain bacteria in the gut may contribute to diarrhea occurrence in yaks.
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Enfermedades de los Bovinos , Diarrea , Heces , Microbioma Gastrointestinal , ARN Ribosómico 16S , Animales , Bovinos , Diarrea/veterinaria , Diarrea/microbiología , Heces/microbiología , Enfermedades de los Bovinos/microbiología , ARN Ribosómico 16S/genética , Bacterias/clasificación , Bacterias/aislamiento & purificación , Bacterias/genéticaRESUMEN
Porcine Epidemic Diarrhea Virus (PEDV) poses a significant threat to neonatal piglets, particularly due to the limited efficacy of existing vaccines and the scarcity of efficacious therapeutic drugs. Gegen Qinlian Decoction (GQD) has been employed for over two millennia in treating infectious diarrhea. Nonetheless, further scrutiny is required to improve the drug's efficacy and elucidate its underlying mechanisms of action. In this study, a modified GQD (MGQD) was developed and demonstrated its capacity to inhibit the replication of PEDV. Animal trials indicated that MGQD effectively alleviated pathological damage in immune tissues and modulated T-lymphocyte subsets. The integration of network analysis with UHPLC-MS/MS facilitated the identification of active ingredients within MGQD and elucidated the molecular mechanisms underlying its therapeutic effects against PEDV infections. In vitro studies revealed that MGQD significantly impeded PEDV proliferation in IPEC-J2 cells, promoting cellular growth via virucidal activity, inhibition of viral attachment, and disruption of viral biosynthesis. Furthermore, MGQD treatment led to increased expression levels of IFN-α, IFN-ß, and IFN-λ3, while concurrently decreasing the expression of TNF-α, thereby enhancing resistance to PEDV infection in IPEC-J2 cells. In conclusion, our findings suggest that MGQD holds promise as a novel antiviral agent for the treatment of PEDV infections.
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Infecciones por Coronavirus , Medicamentos Herbarios Chinos , Farmacología en Red , Virus de la Diarrea Epidémica Porcina , Enfermedades de los Porcinos , Animales , Virus de la Diarrea Epidémica Porcina/efectos de los fármacos , Porcinos , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/química , Infecciones por Coronavirus/tratamiento farmacológico , Infecciones por Coronavirus/virología , Enfermedades de los Porcinos/tratamiento farmacológico , Enfermedades de los Porcinos/virología , Antivirales/farmacología , Replicación Viral/efectos de los fármacos , Línea Celular , Espectrometría de Masas en Tándem , Diarrea/tratamiento farmacológico , Diarrea/virología , Diarrea/veterinaria , Subgrupos de Linfocitos T/metabolismo , Subgrupos de Linfocitos T/efectos de los fármacos , Subgrupos de Linfocitos T/inmunologíaRESUMEN
Clarifying the etiology of diarrhea cases of unknown cause is important in the fight against enteric infections. In this study, we aimed to investigate the role of canine kobuvirus (CaKoV), in cases of diarrhea of unknown origin in dogs. A total 121 swab samples from dogs with diarrhea were collected. Molecular analyses of the samples were performed. For this purpose, after the sequence reaction, a phylogenetic tree was created, and bioinformatics analyses were performed. The prevalence rate of CaKoV in the sampled population was determined as 16.5% (20/121). The presence of parvovirus and coronavirus, which are common viral agents in CaKoV-positive dogs, was determined as 35% (7/20) and 10% (2/20), respectively. The rate of dogs with only CaKoV detected was 65% (13/20). Phylogenetic analysis of CaKoV strains clustered together closely related to reference strains. There are very limited studies on the role of CaKoV in the etiology of diarrhea cases of unknown cause in dogs around the world. So far, only one study has been done on CaKoV in Turkey. In this report which includes molecular characterization and epidemiological data on CaKoV determined the importance of CaKoV in cases of diarrhea of unknown origin. More comprehensive studies are needed to better understand the pathogenesis, epidemiology, and biology of CaKoV and to determine effective strategies to combat it.
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Diarrea , Enfermedades de los Perros , Kobuvirus , Filogenia , Infecciones por Picornaviridae , Perros , Animales , Kobuvirus/genética , Kobuvirus/aislamiento & purificación , Kobuvirus/clasificación , Diarrea/virología , Diarrea/veterinaria , Diarrea/epidemiología , Enfermedades de los Perros/virología , Infecciones por Picornaviridae/veterinaria , Infecciones por Picornaviridae/virología , Infecciones por Picornaviridae/epidemiología , Turquía/epidemiología , Heces/virologíaRESUMEN
Soybean meal is known to be able to cause intestinal damage and dysfunction in early-weaned piglets. However, research on natural compounds that can alleviate these effects is scarce. In this study, the effect of luteolin, a natural flavonoid, on intestinal health of piglets fed on a soybean meal-based diet was explored. A total of eighteen 21-d-old piglets were selected and randomly divided into 3 groups: a negative control group fed with an animal protein-based diet, a positive control group fed with a soybean meal-based diet, and a luteolin group that was fed with the positive control diet supplemented with luteolin. The results suggested that luteolin supplementation significantly increased the average daily gain and average daily feed intake of early-weaned piglets, while effectively reducing the diarrhea incidence. Additionally, luteolin supplementation lowered the levels of soybean antigen-specific immunoglobulin G and immunoglobulin E anitbodies, increased the superoxide dismutase activity in both sera and small intestine mucosa, and enhanced the total antioxidant capacity in sera. Further research found that luteolin supplementation increased the intestinal villi height and decreased the crypt depth, resulting in an increased ratio of villi to crypts. At the same time, it reduced the concentration of serum diamine oxidase, improving intestinal barrier function. Moreover, luteolin significantly decreased the gene expression of Bax and Caspase-3, reducing cell apoptosis in the intestinal mucosa. Luteolin supplementation also increased the abundance of Actinobacteria at the phylum level, reduced the abundance of Prevotella and increased the abundance of Olsenella at the genus level. In conclusion, the supplementation of luteolin to the soybean meal diet was capable of effectively reducing allergic response, enhancing the antioxidant capacity of early-weaned piglets, protecting their intestinal barrier function, inhibiting intestinal mucosal cell apoptosis, and altering the intestinal microbiota structure, therefore promoting intestinal health and improving production performance in early-weaned piglets.
Soybean meal can cause intestinal injury in early-weaned piglets. We wanted to know if luteolin has a protective effect against the intestinal injury caused by soybean meal in piglets. We found that luteolin was capable to alleviate intestinal injury and dysfunction caused by soybean meal in early-weaned piglets, promote their intestinal health, and improve production performance.
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Alimentación Animal , Dieta , Suplementos Dietéticos , Glycine max , Luteolina , Destete , Animales , Luteolina/farmacología , Luteolina/administración & dosificación , Glycine max/química , Porcinos , Alimentación Animal/análisis , Dieta/veterinaria , Suplementos Dietéticos/análisis , Mucosa Intestinal/efectos de los fármacos , Enfermedades de los Porcinos/prevención & control , Antioxidantes/farmacología , Antioxidantes/metabolismo , Intestinos/efectos de los fármacos , Intestinos/patología , Diarrea/veterinaria , Diarrea/prevención & controlRESUMEN
Bovine coronavirus (BCoV) is one of the important causes of diarrhoea in cattle. The virus is responsible for the high fatality rate associated with acute diarrhoea in calves. Rapid and accurate tests need to be conducted to detect the virus and minimise economic losses associated with the disease. Nucleic acid-based detection assays including PCR is an accurate test for detecting pathogens. However, these tests need skilled personnel, time and expensive devices. In this study, we developed a novel assay for the detection of BCoV in clinical cases. This novel assay combined reverse transcription-recombinase polymerase amplification with CRISPR/Cas13 and conducted a rapid visualisation of cleavage activity using a Lateral Flow Device. A conserved sequence of the BCV M gene was used as a target gene and the assays were tested in terms of specificity, sensitivity and time consumption. The result showed the specificity of the assay as 100% with no false positives being detected. Ten copies of the input RNA were enough to detect the virus and perform the assay. It took up to forty minutes for reading the results. Conducted together, the assay should be used as a rapid test to clinically diagnose infectious pathogens including bovine coronavirus. However, the assay needed the RNA to be extracted from the clinical sample in order to detect the virus. Therefore, more studies are needed to optimise the assay to be able to detect the virus in the clinical sample without extracting the RNA.