First Molecular Genotyping of Cryptosporidium felis in Cattle, Iraq.

To the best of our knowledge, this is the first Iraqi study to detect C. felis in cattle by the polymerase chain reaction (PCR) assay and to confirm the local isolates in the National Center for Biotechnology Information (NCBI). Overall, 130 diarrheic calves of different ages and sexes were selected randomly from rural and sub-urban areas in Wasit province (Iraq) from February to April (2021) and subjected to direct collection of fresh fecal samples for DNA extraction and PCR examination. Targeting the heat shock protein 70 (HSP70) gene of C. felis showed that 17.69% of the animals were positive. Findings from clinical examination revealed no significant differences in values of temperature, pulse, or respiratory rates between the positive and negative calves. The association between the positive results and demographic risk factors showed no significant differences in the prevalence rate of infection and risk of exposure to C. felis between the rural and sub-urban areas; however, higher significant values were reported in calves aged 6 months than in calves aged 12 months, as well as in females than in males. Five of the local C. felis isolates were documented under the accession numbers MZ964600.1, MZ964601.1, MZ964602.1, MZ964603.1, and MZ964604.1. Finally, the data presented here provide epidemiological and molecular evidence that the range of C. felis in cattle is wider than expected globally, with a high probability of infection transmission from cattle to humans.

Pathological, Immunological, and Hematological Parameters Associated with Experimental Infection of Citrobacter Freundii in Rabbits.

Citrobacter freundii is one of the most important nosocomial opportunistic pathogens, which causes sepsis, as well as different gross and histopathological lesions in various internal organs in humans and animals, especially dogs and fish. This study aimed to investigate the hematological parameters, immunological responses, and pathological effects of the infection induced by the virulent strain of C. freundii on rabbits. A total of 42 rabbits (local breed; male and female), with a mean weight of 1.5-2 kg, were housed under controlled environmental conditions (20±2°C, 14:10 h light: dark cycle) and allowed ad libitum access to food and water. After two weeks of adaption, the rabbits were divided randomly into three groups of 14 animals per group. Group one (G1) received 3×108 CFU/ml of the virulent isolate (intraperitoneally [IP]) of C. freundii. Group two (G2) was injected subcutaneously (SC) with 3×108 CFU/ml of the virulent strain of C. freundii, while group three was IP injected with phosphate buffer saline and considered a negative control group. Results showed the variable gross pathological effects which included hemorrhage, edema, and congestion of visceral organs. Furthermore, the microscopic lesions showed pneumonia due to inflammatory cells infiltration, mainly neutrophils, macrophages, plasmacytes, and lymphocytes, severe interstitial and intra-alveolar edema, extensive pulmonary hemorrhage, emphysema, and atelectasis. The recorded data from the liver samples revealed hepatitis which was characterized by perivascular and periportal leukocyte cuffing, marked centrilobular with periportal necrosis, extensive hepatic edema, and periportal edema in addition to extensive fibrosis in interlobular septa and periportal fibrosis with severe interstitial hemorrhage. In the kidneys, there were severe renal edema, mixed inflammatory exudation, mainly neutrophils, macrophages, plasmacytes, lymphocytes, fibroblast infiltration in renal parenchyma and renal cortex, extensive renal hemorrhage, edema, as well as fibrosis and severe renal tubular necrosis. In addition, enteritis appeared in the intestine with mucosal edema, especially in lamina propria; moreover, necrosis of entire villi, epithelial necrosis, mucosal and submucosal hemorrhage, and fibrosis were observed. The present study revealed a significant increase in total leukocytes count and the concentration of TNF-α in the infected groups. To the best of the authors' knowledge, this study is considered the first attempt aimed to detect the pathological effects of C. freundii on visceral organs in rabbits. It is concluded that this bacterium could induce a significant pathological, hematological, and immunological changes in the infected animals.