Immunophenotyping of peripheral circulating lymphocytes and serum selenium levels in calves with neonatal diarrhea.

This work aims to: (1) elucidate the immune response exhibited by CD4 + and CD8 + T lymphocyte cells in response to various infectious agents in calves suffering with neonatal diarrhea; and (2) determine and investigate the association between serum selenium levels and T lymphocyte subtypes in neonatal calves afflicted with neonatal diarrhea and infected with various infectious agents. The study encompassed a cohort of 50 calves, encompassing both sexes and various breeds, within the neonatal age range (1-28 days old). Subdivided into distinct groups, the calves were categorized based on the causative agents of neonatal diarrhea, including Rotavirus (n = 10), Cryptosporidium parvum (C.parvum) (n = 10), Coronavirus (n = 5), Rotavirus+C.parvum (n = 5), and a Control group (n = 20). Blood samples were meticulously obtained from the vena jugularis of all animals utilizing specific techniques-8 ml in tubes devoid of anticoagulant and 3 ml in blood collection tubes containing EDTA. Serum selenium levels were analyzed by ICP-MS. Flow Cytometry device was used to determine CD4 + and CD8 +T lymphocyte levels. In this study, although there was no statistically significant difference in serum selenium levels between all study groups, it was found that the selenium level in the control group was not sufficient. CD4 T lymphocyte levels, the rotavirus+C.parvum group exhibited a statistically significant elevation compared to the coronavirus group. Regarding CD8 + T lymphocyte levels, the coronavirus group demonstrated a statistically significant increase when compared to the control group. In intragroup analyses of CD8 + T lymphocyte levels, the coronavirus group exhibited a significant elevation compared to the rotavirus group, C.parvum group, and the C.parvum + Rotavirus group. A significant negative correlation was detected between selenium levels and CD4 + T lymphocytes, while no correlation was found between CD8 + T lymphocytes. Fibrinogen concentration exhibited statistical significance, being higher in the Rotavirus group (p < 0.008) compared to the control group, in the C.parvum group (p < 0.004) compared to the control group, and in the Coronavirus group (p < 0.001) compared to the control group. The leukocyte count demonstrated statistical significance, being higher in the Rotavirus group compared to the control group (p < 0.001), in the Rotavirus+C.parvum group compared to the control group (p < 0.002), and in the Coronavirus group compared to the control group (p < 0.011). In conclusion, the data derived from this study illuminate discernible disparities in CD4 + and CD8 + T lymphocyte immune responses, contingent upon the specific etiological agent associated with neonatal diarrhea. Furthermore, the study underscores the importance of considering selenium deficiency as a relevant factor in calves affected by neonatal diarrhea.

Clinical, pathological and molecular evaluations and CT scan screening of coenurosis (Coenurus cerebralis) in sheep and calves / Avaliação clínica, patológica e molecular e tomografia computadorizada de cenurose (Coenurus cerebralis) em carneiros e bezerros

Abstract The aims of this study were to diagnose coenurosis by means of computerized tomography (CT) scan imaging and molecular characterization of the CO1 gene using the polymerase chain reaction (PCR). Sheep and calves were necropsied, and CT scans on the cephalic region were performed on the animals. Sections of brain tissue infected with parasites were then stained with hematoxylin and eosin for microscopic examination. Material collected from brain cysts was fixed in 70% ethanol. PCR amplification was carried out using the CO1 mitochondrial gene. A total of 60 calves and 80 sheep were examined clinically and, of these, 15 calves and 38 sheep showed signs of depression, with counterclockwise circling movements and altered head carriage. Four sheep and one calf were necropsied, and C. cerebralis cysts were detected in all of them. A hypodense cyst was monitored in the right cerebellar hemisphere on a CT scan on one sheep. A cyst was found in the left frontal lobe on a CT scan on one calf. Microscopically, C. cerebralis cysts were surrounded by a fibrous or epithelial wall that presented necrosis on cerebral sections of both the sheep and the cattle. The CO1-PCR assay yielded a 446 bp band, which was sequenced and phylogenetically analyzed: the results confirmed the presence of T. multiceps. This study reports the first use of CT imaging on naturally infected calves and sheep for diagnosing coenurosis.

Resumo Os objetivos deste estudo foram diagnosticar cenurose por tomografia computadorizada (CT) por imagem de digitalização e caracterização molecular do gene CO1, usando a Reação em Cadeia da Polimerase (PCR). Ovelhas e bezerros foram necropsiados, e uma tomografia computadorizada da região cefálica foi realizada nos animais. Em seguida, cortes microscópicos de cérebro infectado com parasitas foram corados com hematoxilina e eosina e posterior avaliação ao microscópio de luz. Em seguida, o material recolhido de cada cisto cerebral foi fixado em etanol a 70%. A amplificação pela PCR foi realizada utilizando-se o gene mitocondrial CO1. Um total de 60 bezerros e 80 ovelhas foram clinicamente examinados e, desses, 15 bezerros e 38 ovelhas apresentaram sinais de depressão, com movimentos circulares em sentido anti-horário, e desvio da cabeça. Quatro carneiros e uma vitela foram necropsiados, e cistos de C. cerebralis foram detectados nos animais. Um cisto hipodenso foi monitorado no hemisfério cerebelar direito por imagem do CT de um carneiro. O cisto foi encontrado no lobo frontal esquerdo por imagem do CT de um bezerro. Microscopicamente, cistos de C. cerebralis foram envolvidos por uma parede fibrosa ou epitelial, apresentando necrose em ambos os cortes cerebrais de ovinos e de bovinos. O ensaio CO1-PCR produziu uma banda de 446 pb, sequenciado e submetido à filogenia, confirmou ser T. multiceps. Este estudo relata a primeira utilização de imagens de CT em bezerros e ovelhas naturalmente infectados para o diagnóstico de coenurosis.

Clinical, pathological and molecular evaluations and CT scan screening of coenurosis (Coenurus cerebralis) in sheep and calves.

The aims of this study were to diagnose coenurosis by means of computerized tomography (CT) scan imaging and molecular characterization of the CO1 gene using the polymerase chain reaction (PCR). Sheep and calves were necropsied, and CT scans on the cephalic region were performed on the animals. Sections of brain tissue infected with parasites were then stained with hematoxylin and eosin for microscopic examination. Material collected from brain cysts was fixed in 70% ethanol. PCR amplification was carried out using the CO1 mitochondrial gene. A total of 60 calves and 80 sheep were examined clinically and, of these, 15 calves and 38 sheep showed signs of depression, with counterclockwise circling movements and altered head carriage. Four sheep and one calf were necropsied, and C. cerebralis cysts were detected in all of them. A hypodense cyst was monitored in the right cerebellar hemisphere on a CT scan on one sheep. A cyst was found in the left frontal lobe on a CT scan on one calf. Microscopically, C. cerebralis cysts were surrounded by a fibrous or epithelial wall that presented necrosis on cerebral sections of both the sheep and the cattle. The CO1-PCR assay yielded a 446 bp band, which was sequenced and phylogenetically analyzed: the results confirmed the presence of T. multiceps. This study reports the first use of CT imaging on naturally infected calves and sheep for diagnosing coenurosis.