The endemic chub Squalius tenellus (Heckel, 1843) was introduced more than 100 years ago to Lake Blidinje (Bosnia-Herzegovina). Only 1 species of enteric helminth was found in a sample of 35 chubs, the tapeworm Caryophyllaeus brachycollis (Janiszewska, 1953). The paper includes histopathological investigation with identification of innate immune cells involved in host reaction and molecular data allowed correct designation of the cestode species. Of 35 specimens of chub examined, 21 (60%) harboured individuals of C. brachycollis and a total of 1619 tapeworms were counted, the intensity of infection ranged from 1 to 390 worms per fish (46.2 ± 15.3, mean ± s.e.). Histopathological and ultrastructural investigations showed strict contact between the worm's body and the epithelia and increase in the number of mucous cells, rodlet cells among the epithelial cells. Within the tunica propria-submucosa, beneath the site of scolex attachment, numerous neutrophils and mast cells were noticed. This is the first study of the occurrence of C. brachycollis in chub from Lake Blidinje and on the response of the innate immune cells of S. tenellus to this tapeworm. Interestingly, in 3 very heavily infected chubs, perforation of the intestinal wall was documented; this is uncommon among cestodes which use fish as a definitive host.

Non-alcoholic fatty liver disease (NAFLD) is now the leading cause of chronic liver disease, affecting approximately 30% of people worldwide. Histopathology reading of fibrosis patterns is crucial to diagnosing NAFLD. In particular, separating mild from severe stages corresponds to a critical transition as it correlates with clinical outcomes. Deep Learning for digitized histopathology whole-slide images (WSIs) can reduce high inter- and intra-rater variability. We demonstrate a novel solution to score fibrosis severity on a retrospective cohort of 152 Sirius-Red WSIs, with fibrosis stage annotated at slide level by an expert pathologist. We exploit multiple instance learning and multiple-inferences to address the sparsity of pathological signs. We achieved an accuracy of $ 78.98\pm 5.86\% $, an F1 score of $ 77.99\pm 5.64\%, $ and an AUC of $ 0.87\pm 0.06 $. These results set new state-of-the-art benchmarks for this application.

Using morphological and molecular studies, the life cycle of Versteria cuja (Cestoda: Taeniidae) was elucidated, involving subterranean rodents (Ctenomyidae) as intermediate hosts, and the lesser grison, Galictis cuja (Mustelidae), as definitive host. Metacestodes (cysticerci and polycephalic larvae) were found mainly in the liver but also in spleen, pancreas, lungs and small intestine of 2 species of tuco-tucos (Ctenomys spp.) from Chubut, Argentina. Identity of the metacestodes with the adult was based primarily on the number, size and shape of rostellar hooks: 40–48 hooks in 2 rows, particularly small (10–16 μm total length by 6–10 μm wide), composed of handle, blade and guard with characteristic shapes. Genetic analysis (cox1 gen mtDNA) performed on metacestodes from both intermediate hosts corroborated their conspecificity with adults of V. cuja from lesser grisons in the same locality. Histopathological study showed the hepatic parenchyma altered by the presence of cysts containing larvae, each surrounded by a capsule of connective tissue with inflammatory infiltrate, atrophied hepatocytes and an increase of bile ducts. In the lung, in addition to the cysts, dilated alveoli, oedema and hyperaemic blood vessels were observed. This is the first report of a natural life cycle of a Versteria species from South America. It shows strong similarities with that described for a North American zoonotic lineage of Versteria, confirming a close relationship between V. cuja and this North American lineage, as previously demonstrated by molecular studies. Consequently, the zoonotic potential of V. cuja should not be disregarded.

Foot-pad dermatitis (FPD) is an important indicator of animal welfare in turkeys (Meleagris gallopavo). The present study aimed to evaluate the reliability and validity of a commonly used five-point visual score (VC), assessing FPD based on lesion size by implementing histopathological analysis. In 100 turkey feet (20 per scale scored by the VC), the size of both the foot-pad and the alteration were measured. Subsequently, a histopathological analysis was performed, examining the occurrence and severity grade of different parameters. The study addressed three main goals: (i) examining the reliability of the scoring system concerning the evaluated size of FPD; (ii) assessing histopathological parameters to analyse patterns, reflecting the categories of the scoring system; and (iii) finding threshold values to avoid the occurrence of ulcerations. The study found good observer reliabilities for the VC, but measuring the relative size of alterations resulted in divergent values according to the original specifications With regard to the histopathological parameters, no clear patterns were found in the respective VC scoring levels. However, ulcerations revealed a significant effect on the size of the alteration, showing a greater grade of severity with increasing lesion size. Regarding the occurrence of ulcerations, optimum threshold values could be identified even in very small lesions. This study helps contribute to a better understanding of the pathophysiology of FPD. It also raises the question as to whether, in light of animal welfare concerns, threshold values of visual systems should be adjusted to avoid ulcerations, considering the outcome of our histopathological assessment.

Neonatal mortality in puppies is a problem frequently encountered by dog breeders. Often, only postmortem examination allows diagnosis and implementation of measures to save the rest of the litter. This article presents the key steps of the postmortem examination, namely, autopsy, histopathology, bacteriology, molecular identification of pathogens, and coproscopy. Sampling, samples' conservation, and interpretation of the obtained results are presented as well as their relative importance for the final diagnosis. Finally, examples of the most frequent syndromes observed under postmortem examination in canine newborns, together with the results from a complementary analysis looking for infectious agents responsible for death, are discussed.

The current study was aimed to evaluate the effects of variable doses of the weedicide glyphosate on the ileal (the final section of the small intestine) structure of rats of both sexes, using histological, histochemical, and ultrastructural methods. Forty animals were classified into four groups of 10 animals per group (five males and five females). The first group acted as a control, and the remaining groups were treated with glyphosate-Roundup® 25, 50, and 100 mg/kg body weight daily for 15 days. The results indicated extinct histopathological changes manifested in the deformation of villi, foci of leukocytic infiltration in the core of villi, and hyperplasia of goblet cells. Histochemical examination (Alcian blue and Periodic acid–Schiff stain) revealed a strong positive reaction of goblet cells and an increase in their number in all treated groups. In addition, the immunohistochemical investigation revealed the immunoreactivity of matrix metalloproteinase-9 expression. Furthermore, electron microscopic alternations were represented by the deformation of nuclei, destruction of microvilli, and deposition of lipid droplets. Collectively, the present findings indicate that treatment with glyphosate results in extensive morphological alternations to the ileal structure of rats of both sexes and that female rats are more affected than male rats are.

Ectopic calcification or pathological biomineralization correlates with morbidity and mortality from cardiovascular diseases. Aortas with atherosclerotic lesions and biomineralization were selected for the study. Thirty samples of mineralized abdominal aortas (group M) were examined by histology. Depending on the calcifications size, samples were separated into group M1 (macroscopic calcifications) and M2 (microscopic calcifications). Each group consists of 15 samples. Calcification 2 mm or less were considered as microscopic, >2 mm—macroscopic. Thirty samples of aortic tissue without biomineralization (group C) were used as a control group. Aortic tissue was examined by macroscopic description, histology, histochemistry, immunohistochemistry (IHC), scanning electron microscopy (SEM) with microanalysis, and transmission electron microscopy (TEM). The results of IHC showed the involvement of OPN in the formation and development of pathological biomineralization, but the obvious role of OPN in the differentiation of macro- and microcalcifications of atherosclerotic aorta was not revealed. SEM with X-ray microanalysis confirmed that the biomineral part of the aortic samples of the M1 group consisted mainly of apatites, which correspond to previous studies. The Ca/P ratio was less in the M2 group than in the M1 group. It means that microcalcifications can be formed by more defective (immature) hydroxyapatite.

This study describes changes in haematological parameters, cytokine profile, histopathology and cortisol levels in Swiss mice experimentally infected with Angiostrongylus costaricensis. Twenty-eight Swiss mice were divided into two groups (G1 and G2) of 14 animals each. In each group, eight animals were infected orally with ten third-stage larvae of A. costaricensis and six were used as a control group. The mice of groups G1 and G2 were sacrificed 14 and 24 days after infection, respectively. Samples were collected for histopathological and haematological analyses and determination of the cytokine profile and cortisol levels. Granulomatous reaction, eosinophilic infiltrate and vasculitis in the intestinal tract, pancreas, liver and spleen were observed with varying intensity in infected animals. Our results showed that the mice developed normocytic and hypochromic anaemia, and that the histopathological lesions caused by the experimental infection influenced increases in cortisol, neutrophil and monocyte levels. In addition to this, we detected increased interleukin-6 and tumour necrosis factor alpha levels in the infected animals.

The California kingsnake (Lampropeltis californiae), native to North America, is a significant threat to the conservation of endemic species in the Spanish Macaronesian island of Gran Canaria. However, its role disseminating potential invasive parasites, such as zoonotic pentastomids, has not been proven. Among its parasitic fauna, only protistans have been documented, in contrast to other Lampropeltis spp., which are known to carry pentastomids. Thus, a parasitological study is urgently required. Between 2016 and 2018, a total of 108 snakes were necropsied and stool samples examined. A single snake was infested with Ophionyssus natricis, and another individual with Serpentirhabdias sp. Only this latter snake presented gross lesions, characterized by granulomatous pneumonia. No Pentastomida were found. By contrast, almost the entire population (98.5%) was infested with larval helminths (three different nematode and two cestode species), characterized by granulomatous gastrointestinal serositis. This suggests the snake poses a ‘dead end’ host for local parasites. Based on these findings, snakes in Gran Canaria carry potential zoonotic mites, which along with Serpentirhabdias sp. could represent a threat to endemic lizards. The presence of metazoan parasites and their lesions are reported for the first time in the California kingsnake.

Tramadol is used worldwide and is listed in many medical guidelines to treat both acute and chronic pains. There is a growing evidence of abuse of tramadol in some African and West Asian countries. Tramadol has some side effects. The present study designed to follow up the treatment of the cellular responses which might be induced in the kidney of tramadol mice. Treated mice received daily injection of tramadol dose (125 μg/100 g b.wt) for 20 and 40 days. Other mice received tramadol for 40 days and then were divided into three groups: the first received distilled water, the second received Lagenaria siceraria, and the third received melatonin daily for 40 days. Both the daily injection of tramadol for 20 and 40 days resulted in radical, extensive, and severe alterations in the normal histological architecture of the kidney. Treatment with Lagenaria siceraria or melatonin after tramadol administration for a long-term, markedly changed the collagen content and other chemical components, that may reach nearly normal levels. Such findings propose that although tramadol has many cytological and histopathological side effects on the kidneys of male mice, the treatments via Lagenaria siceraria and melatonin have effective therapeutic impacts on the tramadol side effects.

Understanding of a disease and its treatment relies heavily on visualizing cells, tissues, and organs, making micro- and macro-imaging modalities integral to the biomedical community. Macro imaging modalities such as magnetic resonance imaging (MRI), computed tomography (CT), and ultrasound are still the mainstay of whole-body imaging because of their non-invasive nature and live image feed. However, they fail to deliver cellular details required to understand a disease or effectiveness of a treatment. On the opposite end of the spectrum, benchtop microscopes and histology can deliver excellent cellular resolution, allowing one to visualize cellular and intracellular interactions to better understand a disease, its progression and/or the outcome of therapies. However, to look at the cells, the tissue needs to be resected, fixed, processed, and stained before imaging. This whole process of visual confirmation may take hours to days. Deciding on these two imaging modalities always involves a compromise, either in time, resolution, or invasiveness. Confocal laser endomicroscopy (CLE) fills a unique niche between these macro- and micro-imaging modalities as it facilitates non-invasive to minimally invasive in vivo image acquisition in real time. In this two-part series we first examine the equipment available for CLE, and in the second part we will explore several applications of the technology.

Fasciolosis is a food-borne disease that causes great distress to a range of hosts, including humans. The objectives of this study were to (1) evaluate the liver damage and carcass weight of cattle naturally infected with Fasciola hepatica from the state of Rio Grande do Sul (RS), Brazil, and to (2) determine the distribution of adult flukes in 12,236 cattle liver from RS. The data from these experiments were used to calculate the overall economic loss due to F. hepatica infection. Eighteen adult Polled Hereford cows were divided into a triclabendazole (TbG) and a F. hepatica-positive group (FhG). For Experiment 1, a generalized linear mixed model revealed a statistical difference in carcass weight (49.8 kg) between TbG and FhG. The Monte Carlo analysis also revealed that the animals’ weight differences were due to the disease. For Experiment 2, the prevalence of infected livers was above 16% (1904/12,236), mostly (20.1%) from the south-west region of RS. The Susceptible-infected-recovered (SIR) epidemic model revealed the evolution of the infection using a high infectivity and low recovery rate. Other distinctive scenarios that occur in RS were also established with different rates of infectivity. The economic assessment showed a potential loss of US$45 million to the beef cattle industry of RS, with an overall State cost of US$90.3 million. These novel findings reveal the importance of fasciolosis infection, which can cause a significant health condition and poor animal welfare.

Abdominal angiostrongyliasis is caused by Angiostrongylus costaricensis, the definitive and intermediate hosts of which are wild rodents and terrestrial molluscs, respectively. Humans are accidental hosts and can be infected by ingesting the third-stage (infective) larvae (L3). It remains unclear whether the number of L3 inoculated is related to lesion severity. Our aim was to analyse histopathological alterations in Swiss mice infected with different doses of A. costaricensis. Thirty-two mice were randomly divided into four groups (n = 8/group): uninfected, control mice; mice infected with a low dose (five L3); mice infected with an intermediate dose (15 L3); and mice infected with a high dose (30 L3). The frequency of intestinal thrombi, splenitis, eggs/larvae, hepatic infarction and acute pancreatitis differed among the groups, the last being considered a significant finding. We conclude that different infective doses alter the histopathological aspects of the infection in Swiss mice, those aspects being more pronounced at medium and high doses, with no effect on the development of the disease. This experimental model shows that the parasite life cycle can be maintained in Swiss mice through the inoculation of a low dose (five L3).

Recent data suggest that organic broilers often score worse on footpad lesions than conventional broilers but also that the current scoring of organic broiler feet may be misleading. In order to characterise footpad lesions in organic broilers, this study assessed and compared footpad lesions in a sample of 2987 conventional and 3578 organic broiler feet obtained from a large Danish abattoir during summer and winter. The feet were scored according to two scoring systems: the modified Danish surveillance scoring system and a histopathology-based new scoring system specifically developed to target the ability to differentiate between broiler feet with hyperkeratosis and ulcers. For both systems, all broiler feet with visible lesions were cross-sectionally incised. Significant differences between the two production systems were found for both scoring systems (χ2 = 710; P < 0.001 and χ2 = 247; P < 0.001 for the new and the surveillance systems, respectively), showing that a larger proportion of the organic feet compared to conventional feet – summer and winter – exhibited signs of hyperkeratosis. In addition, a smaller fraction of the organic feet than of the conventional feet were given the outermost scores, that is, normal or ulcerated; 13.4% v. 25.3% broiler feet were given score 0 for organic v. conventional production systems, respectively (χ2 = 152; P < 0.001), and 18.4% v. 23.8% feet were given score 4 for organic v. conventional production systems, respectively (χ2 = 308; P < 0.001). Thus, the results suggest that surveillance scoring systems such as the one used in Denmark are useful for the examination of footpad lesions in broilers from both types of production systems. However, the results have also raised attention to a typical characteristic of the feet of organic broilers, that is, profound hyperkeratosis, which may underlie potential misclassifications in surveillance scoring systems like the one used in Denmark. Among the possible solutions to this challenge to the correctness and fairness of the scoring system are improved procedures (such as mandatory incision), training of technicians and calibration of results (especially for the organic footpads).

The present study determined the prevalence of hydatid cysts in different organs of slaughtered hilly ‘Gaddi’ breed small ruminants—sheep (n = 230) and goats (n = 197)—in Kangra Valley of the north-western Himalayas, India. Hydatid cysts were found in 12.2% (n = 28) of sheep and 10.7% (n = 21) of goats. Pulmonary echinococcosis was more prevalent in slaughtered sheep and goats (sheep 56.36%; goats 62.90%) than hepatic echinococcosis (sheep 43.64%; goats 37.10%). Fertility rates were higher in hepatic (81.25%) and pulmonary cysts of sheep (83.87%) compared to goats. Molecular identification and genotypic characterization of Echinococcus granulosus isolates were based on mitochondrial cytochrome oxidase 1 gene (mtCO1). The genotypic characterization identified the isolated strain to be closely related to the G7 genotype. Histopathological examination revealed a thick coat of granulation tissue, causing fibrosis and inflammatory reaction composed of fibroblasts and mononuclear cells around the cysts. In the liver, hepato-cellular degeneration was prominent at the periphery of the cysts. The present study highlights the molecular confirmation and phylogenetic analysis of E. granulosus isolates with the prevalence of hydatidosis in a naïve host species and in an unexplored region. The findings are of significant medical and veterinary importance regarding development of control measures to check dissemination of hydatidosis.