One health surveillance strategy for coronaviruses in Italian wildlife.

The recent reinforcement of CoV surveillance in animals fuelled by the COVID-19 pandemic provided increasing evidence that mammals other than bats might hide further diversity and play critical roles in human infectious diseases. This work describes the results of a two-year survey carried out in Italy with the double objective of uncovering CoV diversity associated with wildlife and of excluding the establishment of a reservoir for SARS-CoV-2 in particularly susceptible or exposed species. The survey targeted hosts from five different orders and was harmonised across the country in terms of sample size, target tissues, and molecular test. Results showed the circulation of 8 CoV species in 13 hosts out of the 42 screened. Coronaviruses were either typical of the host species/genus or normally associated with their domestic counterpart. Two novel viruses likely belonging to a novel CoV genus were found in mustelids. All samples were negative for SARS-CoV-2, with minimum detectable prevalence ranging between 0.49% and 4.78% in the 13 species reaching our threshold sample size of 59 individuals. Considering that within-species transmission in white-tailed deer resulted in raising the prevalence from 5% to 81% within a few months, this result would exclude a sustained cycle after spillback in the tested species.

Main causes of death of free-ranging bats in Turin province (North-Western Italy): gross and histological findings and emergent virus surveillance.

BACKGROUND: Bats are recognized as reservoir species for multiple viruses. However, little is known on bats' health and mortality. Thus, this study aimed to investigate the main causes of death of bats from Turin province (North-western Italy) and to describe gross and histopathological lesions potentially associated with the presence of selected bat viruses. RESULTS: A total of 71 bats belonging to 9 different species of the families Vespertilionidae and Molossidae were necropsied and samples of the main organs were submitted to histopathological examination. Also, aliquots of the small intestine, liver, spleen, lung, and brain were collected and submitted to biomolecular investigation for the identification of Coronaviridae, Poxviridae, Reoviridae (Mammalian orthoreovirus species), Rhabdoviridae (Vaprio ledantevirus and Lyssavirus species) and Kobuvirus. The majority of bats died from traumatic lesions due to unknown trauma or predation (n = 40/71, 56.3%), followed by emaciation (n = 13/71,18.3%). The main observed gross lesions were patagium and skin lesions (n = 23/71, 32.4%), forelimbs fractures (n = 15/71, 21.1%) and gastric distension (n = 10/71,14.1%). Histologically, the main lesions consisted of lymphoplasmacytic pneumonia (n = 24/71, 33.8%), skin/patagium dermatitis (n = 23/71, 32.4%), liver steatosis and hepatitis (n = 12, 16.9%), and white pulp depletion in the spleen (n = 7/71, 9.8%). Regarding emergent bat viruses, only poxvirus (n = 2, 2.8%) and orthoreovirus (n = 12/71, 16.9%) were detected in a low percentage of bats. CONCLUSIONS: Trauma is the main lesion observed in bats collected in Turin province (North-western Italy) associated with forelimb fractures and the detected viral positivity rate seems to suggest that they did not represent a threat for human health.

Gut health of horses: effects of high fibre vs high starch diet on histological and morphometrical parameters.

BACKGROUND: The conventional feeding management of horses is still characterized by high starch and low fibre diets, which can negatively affect horse's gastrointestinal health. Thus, the aim of this study was to compare the effects of a high-starch (HS) vs. a high-fibre (HF) diet on gut health in horses. A total of 19 Bardigiano horses destined for slaughter and aged 14.3 ± 0.7 months were randomly allotted to two dietary groups: HS (5 fillies and 4 colts,) and HF group (7 fillies and 3 colts). They received the same first-cut meadow hay but different complementary feeds for 72 days: HS group was fed 8 kg/animal/day of a starch-rich complementary feed while HF group was fed 3.5 kg/animal/day of a fibre-rich complementary feed. At slaughter, stomachs were separated and washed for the evaluation of the glandular and squamous regions. Also, duodenum, jejunum, ileum, apex of the caecum, sternal flexure, pelvic flexure, right dorsal colon, rectum and liver were excised and submitted to histomorphometrical evaluation. RESULTS: The glandular region of HS group presented more severe gastric mucosa lesions compared to the HF group (P = 0.006). Moreover, a statistical tendency (P = 0.060) was found for the squamous region, presenting a higher score in HS than HF diet. Regarding morphometry, in jejunum villus height to crypt depth (Cd) ratio was influenced by sex, being greater in males than in females (P = 0.037) while in ileum Cd depended on interaction between sex and diet, being greater in males of HS group (P = 0.029). Moreover, in the duodenum and right dorsal colon the severity of the inflammation depended on sex (P = 0.024 and 0.050), being greater in females than in males. On the contrary, in the jejunum and in the pelvic flexure, inflammation was influenced by diet, being more severe in HS than in HF group (P = 0.024 and 0.052). CONCLUSIONS: These results suggested that HS diet provoked more severe mucosa lesions in the glandular region of the stomach and a higher inflammation both in the jejunum and pelvic flexure. The present study can represent a starting point for further investigations on gut health in horses.

A high-starch vs. high-fibre diet: effects on the gut environment of the different intestinal compartments of the horse digestive tract.

BACKGROUND: Horses are often fed high amounts of starch in their diets despite the well-established benefits of a fibre-based diet to promote gut health and animal welfare. The aim of the present study was to compare the effects of two different diets - one based on high amounts of starch (HS) vs. one base on high amounts of fibre (HF) - on specific parameters of the gut environment across different intestinal compartments of the horse digestive tract. To this end differences in the gastrointestinal environment between HS vs. HF fed horses were assessed in terms of dry matter, organic matter and ash content; the particle size distribution and volatile fatty acid composition were also investigated. RESULTS: Nineteen Bardigiano horses of 14.3 ± 0.7 months of age and destined to slaughter were divided into two group pens - one fed with high amounts of starch (HS; n = 9; 43% hay plus 57% starch-rich pelleted feed); vs. fed with high amounts of fibre (HF; n = 10; 70% hay plus 30% fibre-rich pelleted feed). Horses fed HS diet presented a higher dry matter content in the right dorsal colon. Moreover, they showed a higher organic matter and ash content in the sternal flexure, pelvic flexure, right dorsal colon and rectum. In these latter intestinal compartments, horses fed a HS diet also showed a higher proportion of particles retained on an 8 mm sieve and a higher proportion of particles that washed through the finest sieve (< 1 mm). Moreover, the total amounts of volatile fatty acids as well as valeric acid were found to be significantly higher in horses fed the HS vs. HF diet. CONCLUSIONS: A high-starch diet causes significant changes in the horse gut environment. We observed an increase in the dry matter content in the right dorsal colon, as well as reduced particle sizes and an increase in the production of valeric acid in all the gut compartments studied. High-starch diets should be avoided in favour of fibre-based diets with the goal of safeguarding gut health in horses.

Pulmonary fibrosis in a dog as a sequela of infection with Severe Acute Respiratory Syndrome Coronavirus 2? A case report.

BACKGROUND: Interstitial lung disease is a heterogeneous group of conditions characterized by severe radiographic changes and clinicopathological findings. However, in the vast majority of cases, the cause remains unknown. CASE DESCRIPTION: In the present study, we reported the clinical case of a 3 years old female Bull Terrier presented in October 2020 to the Advanced Diagnostic Imaging Department of the Turin Veterinary Teaching Hospital with a progressive pulmonary illness characterized by dyspnea, exercise intolerance, and a diffuse and severe pulmonary interstitial pattern at imaging investigations. Considering the clinical findings, the dog was included in a serological survey for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection in companion animals, showing positive results. Due to the further clinical worsening, the owners opted for euthanasia. At necroscopy, dog showed severe and chronic bronchopneumonia compatible with a Canine Idiopathic Pulmonary Fibrosis and with serological features linked to a SARS-CoV-2 infection. CONCLUSIONS: The comparison of these lesions with those reported in humans affected by Coronavirus Disease 2019 (COVID-19) supports the hypothesis that these findings may be attributable to the post-acute sequelae of SARS-CoV-2 infection in a dog with breed predisposition to Canine Idiopathic Pulmonary Fibrosis (CIPF), although direct evidence of SARS-CoV-2 by molecular or antigenic approaches remained unsolved.

Teleost fish: a new spontaneous model for the study of Lambl's excrescences.

Lambl's excrescences (LE) are fibrous strands typically occurring at coaptation lines of cardiac valves. Although they have been widely reported in human medicine, information on LE occurrence in veterinary medicine is still scarce. The present study aimed to investigate the presence of LE in different freshwater fish and in swordfish. A total of 185 hearts were collected from different fish species, and samples of different cardiac areas (sinus venosus, atrial wall with sinoatrial valves, ventricular wall with atrioventricular valves and bulb with bulboventricular valves) were submitted to histopathological evaluation. LE were detected in 6 of 103 freshwater fish (5.8%) and 19 of 82 swordfish (23.2%). LE developed in atrioventricular, sinoatrial and bulboventricular valves. All affected valves also showed endocardiosis. Based on the results of the present work, teleost fish, specifically swordfish, could be proposed as a novel spontaneous model for the study of LE pathogenesis.

The effect of dietary supplementation with globin and spray-dried porcine plasma on performance, digestibility and histomorphological traits in broiler chickens.

This study evaluated the effects of globin and spray-dried porcine plasma (SDPP) on growth performance, digestibility, nitrogen retention, energy retention efficiency (ERE) and intestinal morphology of broiler chickens. A total of 336-day-old male broiler chickens were reared from 1 to 40 days of age and fed 3 diets (8 replicates/diet, 14 birds/replicate) during 3 feeding phases: starter (1-12 days), grower (12-25 days) and finisher (25-40 days). Isonitrogenous diets were formulated by replacing gluten protein isolate contained in the control diet (C diet) with 2% (starter) or 1% (grower and finisher) spray-dried porcine plasma in the plasma diet (SDPP diet). The globin diet (G diet) was obtained by adding globin on the top of C diet at a dose of 0.08% for the whole rearing period. Total tract apparent digestibility (aD), nitrogen retention and ERE were assessed during the three growing phases. At 12  and 40 days of age, one bird per pen was slaughtered to sample gut, liver, spleen and bursa of Fabricius for histomorphological investigations. The SDPP diet increased body weights of chickens at 12 (+60 g; p < .001), 25 (+101 g; p < .001) and 40 days (+130 g; p = .018) of age compared to C and G diets. Also SDPP improved crude protein aD (+9.7%) and ERE (+12.3%) during the starter phase (p < .001). Dietary globin and SDPP inclusion did not affect either the gut morphology or the histopathological findings in birds at 12 and 40 days of age, despite a numerical (+6.90% and +7.40% respectively) villus height improvement in the SDPP group. Overall, these results confirm that dietary supplementation with SDPP and, to a lesser extent, with globin can improve growth performance and dietary protein and energy utilization in broiler chickens without effect on gut functionality.

Black soldier fly and yellow mealworm live larvae for broiler chickens: Effects on bird performance and health status.

The commercial broiler chicken strains are the result of successful selection programmes. Most of the problems related to welfare arise from the high growth rate and body weight. The use of environmental enrichments in intensive farming could have a positive effect on birds by increasing animal welfare. The aim of the study was to evaluate the effects of Hermetia illucens (HI) and Tenebrio molitor (TM) live larvae in the diets of broiler chickens on growth performance, carcass yield and health status. A total of 180 four-day-old male broiler chickens (Ross 308) were randomly allotted to 18 pens. Each pen was assigned to one of the three dietary treatments (6 replicates/treatment, 10 birds/replicate) as follows: (i) control diet (C): commercial feed (two feeding phases: starter [4-11 days] and grower [12-38 days]), (ii) HI: C + 5% of the expected daily feed intake (DFI) HI live larvae (calculated on dry matter [DM]) and (iii) TM: C + 5% of DFI TM live larvae (DM). At 39 days of age, birds were slaughtered. Growth performance parameters were overall not affected by dietary treatments, except for the grower phase feed conversion ratio (FCR) and the overall FCR being better in the TM broilers than the others (p < 0.01). No differences were observed for slaughtering performance and haematological and serum parameters, except for the spleen relative weight being higher (p < 0.01) in the birds administered with larvae when compared to the C group. Gut morphometric indexes and histopathological alterations were not influenced by insect larvae administration. In conclusion, the administration in limited quantities of HI and TM live larvae as environmental enrichment has no negative effects on broiler chicken growth performance and health status. A behavioural study could confirm that live insect larvae represent a novel natural environmental enrichment in broiler farming.

Cistus salviifolius Toxicity in Cattle.

Toxicity related to consumption of Cistus sp. pl. has been described in ruminants in some countries. This report describes the clinical and pathological findings of Cistus salviifolius toxicosis in 3 beef cattle herds located in 2 different areas of Sicily, Italy. Outbreaks were observed after grazing in poor winter pasture where C. salviifolius was abundant. Mean morbidity and mortality were 29% and 21%, respectively. Most of the affected animals (6 to 36 months old) showed anorexia, weight loss, and pollakiuria culminating in recumbency and death. Occasionally, abortion and neurological signs were observed. In animals with acute signs, there was a moderate decrease of sodium and chloride concentrations in serum. Animals with chronic signs showed an increase of serum urea, creatine phosphokinase (CPK), aspartate transaminase (AST), lactate dehydrogenase (LDH), and phosphorus and a decrease in total serum protein, calcium, chloride, and magnesium concentrations. Moderate anemia and slight neutropenia, lymphocytosis, and eosinophilia were detected in all groups. At necropsy, the main lesion was severe distention of the urinary bladder with turbid hemorrhagic urine and crystalluria. Histologically, chronic cystitis, interstitial nephritis, eosinophilic enteritis, and nonsuppurative necrotizing hepatitis were observed. To our knowledge, this is the first report of C. salviifolius toxicosis in cattle in Italy.

Validation of Thiosemicarbazone Compounds as P-Glycoprotein Inhibitors in Human Primary Brain-Blood Barrier and Glioblastoma Stem Cells.

P-glycoprotein (Pgp) is highly expressed on blood-brain barrier (BBB) and glioblastoma (GB) cells, particularly on cancer stem cells (SC). Pgp recognizes a broad spectrum of substrates, limiting the therapeutic efficacy of several chemotherapeutic drugs in eradicating GB SC. Finding effective and safe inhibitors of Pgp that improve drug delivery across the BBB and target GB SC is open to investigation. We previously identified a series of thiosemicarbazone compounds that inhibit Pgp with an EC50 in the nanomolar range, and herein, we investigate the efficacy of three of them in bypassing Pgp-mediated drug efflux in primary human BBB and GB cells. At 10 nM, the compounds were not cytotoxic for the brain microvascular endothelial hCMEC/D3 cell line, but they markedly enhanced the permeability of the Pgp-substrate doxorubicin through the BBB. Thiosemicarbazone derivatives increased doxorubicin uptake in GB, with greater effects in the Pgp-rich SC clones than in the differentiated clones derived from the same tumor. All compounds increased intratumor doxorubicin accumulation and consequent toxicity in GB growing under competent BBB, producing significant killing of GB SC. The compounds crossed the BBB monolayer. The most stable derivative, 10a, had a half-life in serum of 4.2 h. The coadministration of doxorubicin plus 10a significantly reduced the growth of orthotopic GB-SC xenografts, without eliciting toxic side effects. Our work suggests that the thiosemicarbazone compounds are able to transform doxorubicin, a prototype BBB-impermeable drug, into a BBB-permeable drug. Bypassing Pgp-mediated drug efflux in both BBB and GB SC, thiosemicarbazones might increase the success of chemotherapy in targeting GB SC, which represent the most aggressive and difficult components to eradicate.

Lesions Consistent with Tuberculous Spondylitis in Domestic and Wild Swine and Their Potential Use as a Model for Pott Disease in Humans.

During a routine abattoir inspection of pig carcasses and control activities of hunted wildlife in 2013, 118 large white pigs Sus scrofa domesticus, 474 Nebrodi black pigs and 135 wild boars Sus scrofa scrofa were submitted to anatomopathological examination to evaluate the presence of tuberculosis-like lesions. Localized and generalized granulomatous lesions were detected with a prevalence of about 25% in large white pigs, 13% in Nebrodi black pigs and 8.15% in wild boars. Localized lesions involved mainly the submandibular lymph nodes, but when the disease was spread throughout the body, the inner organs and, also, in some cases, udders and/or bones were injured. The highest prevalence of generalized lesions (15/30) was observed in large white pigs, the only ones in which tuberculous granuloma affected also the spine. The bovine tuberculous spondylitis cases observed showed some similarities with Pott disease in humans regarding aspect and localization of lesions and age of the affected animals.Tissue samples of the positive animals were collected and submitted to bacteriological analysis, and the bone samples were also subjected to histological and immunohistochemistry analysis. M. bovis was isolated in all the analysed samples, and the granuloma encapsulation was found often incomplete indicating that the disease was in an active phase. The presence of lesions associated to tuberculous spondylitis in pigs suggests the possibility to use this animal species as model for the study of Pott disease in humans.

Modulation of intestinal microbiota, morphology and mucin composition by dietary insect meal inclusion in free-range chickens.

BACKGROUND: Gut health in poultry depends on the balance between the host, intestinal microbiota, intestinal microscopic features and diet. The effects of insect meal (a promising alternative protein source for poultry feed) on chicken gut morphology have recently been reported, but no data about intestinal microbiota and mucin composition modulation are available. The present study evaluated the effects of dietary Tenebrio molitor (TM) meal inclusion on gut health of free-range chickens by intestinal microbiota, morphology and mucin composition characterization. RESULTS: One hundred forty female medium-growing hybrids were divided into 2 dietary treatments (control feed [C] and 7.5% TM inclusion, with 5 replicate pens/treatment and 14 birds/pen) and slaughtered at 97 days of age (2 birds/pen for a total of 10 chickens/diet). The gut microbiota assessment on cecal content samples by 16S rRNA amplicon based sequencing showed higher alpha (Shannon, P < 0.05) and beta (Adonis and ANOSIM, P < 0.001) diversity in birds fed TM diet than C. In comparison with C group, TM birds displayed significant increase and decrease, respectively, of the relative abundances of Firmicutes and Bacteroidetes phyla, with higher Firmicutes:Bacteroidetes ratios (False Discovery Rate [FDR] < 0.05). The relative abundance of Clostridium, Oscillospira, Ruminococcus, Coprococcus and Sutterella genera was higher in TM chickens than C (FDR < 0.05). On the contrary, TM birds displayed significant decrease of the relative abundance of Bacteroides genus compared to the C group (FDR < 0.05). Gut morphology evaluation by morphometric analysis on small intestine revealed similar villus height, crypt depth and villus height to crypt depth ratio between C and TM birds. Characterization of gut mucin composition by periodic-acid Schiff, Alcian Blue pH 2.5 and high iron diamine staining on small and large intestine showed unaffected mucin staining intensity in TM chickens when compared to C group. CONCLUSIONS: Dietary TM meal inclusion may positively modulate the gut microbiota of the free-range chickens without influencing the intestinal morphology and mucin composition. Since the rapid growth of chickens directly depends on morphological and functional integrity of the digestive tract, the gut health assessment by a post mortem multidisciplinary approach appears to be fundamental.

Solid Lipid Nanoparticles Carrying Temozolomide for Melanoma Treatment. Preliminary In Vitro and In Vivo Studies.

AIM: To develop an innovative delivery system for temozolomide (TMZ) in solid lipid nanoparticles (SLN), which has been preliminarily investigated for the treatment of melanoma. MATERIALS AND METHODS: SLN-TMZ was obtained through fatty acid coacervation. Its pharmacological effects were assessed and compared with free TMZ in in vitro and in vivo models of melanoma and glioblastoma. RESULTS: Compared to the standard free TMZ, SLN-TMZ exerted larger effects, when cell proliferation of melanoma cells, and neoangiogeneis were evaluated. SLN-TMZ also inhibited growth and vascularization of B16-F10 melanoma in C57/BL6 mice, without apparent toxic effects. CONCLUSION: SLN could be a promising strategy for the delivery of TMZ, allowing an increased stability of the drug and thereby its employment in the treatment of aggressive malignacies.

Antimicrobial Activity of Lactoferrin-Related Peptides and Applications in Human and Veterinary Medicine.

Antimicrobial peptides (AMPs) represent a vast array of molecules produced by virtually all living organisms as natural barriers against infection. Among AMP sources, an interesting class regards the food-derived bioactive agents. The whey protein lactoferrin (Lf) is an iron-binding glycoprotein that plays a significant role in the innate immune system, and is considered as an important host defense molecule. In search for novel antimicrobial agents, Lf offers a new source with potential pharmaceutical applications. The Lf-derived peptides Lf(1-11), lactoferricin (Lfcin) and lactoferrampin exhibit interesting and more potent antimicrobial actions than intact protein. Particularly, Lfcin has demonstrated strong antibacterial, anti-fungal and antiparasitic activity with promising applications both in human and veterinary diseases (from ocular infections to osteo-articular, gastrointestinal and dermatological diseases).

Severe meningeal fibrinoid vasculitis associated with Theileria taurotragi infection in two short-horned Zebu cattle.

The Authors describe a severe vasculitis with fibrinoid necrosis of the meningeal arteries observed in two brains of indigenous short-horn zebu (Bos indicus) cattle, with bovine cerebral theileriosis (BCT) caused by a tick-transmitted hemoprotozoan, Theileria taurotragi, from Northern Tanzania. In the Author's opinion, the role of T. taurotragi infection in the angiocentric and angiodestructive detected features remains to be evaluated. A possible immunopathologic cancerous mechanism, secondary to the lymphoid deregulation, could be involved. This report suggests further studies to better characterize the lymphoid cell involvement in the pathogenesis of the meningeal vascular lesions by T. taurotragi.

"Ormilo disease" a disorder of zebu cattle in Tanzania: bovine cerebral theileriosis or new protozoan disease?

"Ormilo" disease is a neurological disorder of cattle described by Maasai herders in Tanzania. It is attributed to infection by Theileria species, although no detailed data are available in the literature. The authors describe the macroscopical and histological changes observed in 30 brains of indigenous short-horn zebu cattle from Northern Tanzania, aged 2-9 years, with the characteristic neurological signs of "Ormilo". Moreover, the ultrastructural details observed in 14 selected brain samples were reported. Areas of congestion and hemorrhages, associated with the obstruction of the cerebral vessels with large numbers of parasitized lymphoid cells, were observed. Electron microscopy showed the presence of intralymphocytic parasites morphologically comparable to flagellated protozoa, not previously described in the lymphoid cells of cattle, but only reported during the sexual stages within the vector. Theileria taurotragi was detected by polymerase chain reaction (PCR) and reverse line blot (RLB) in nine samples. The authors hypothesize that the parasite detected by electron microscopy could be a strain of a Theileria endemic to this region till now not investigated, having an intralymphocytic phase and being associated with other Theileria spp. infestation. Further studies are needed to better understand the etiology of "Ormilo" disease and to characterize the morphology of the observed parasite, clarifying its role in the disease in Tanzania.

Degenerative myelopathy in German Shepherd Dog: comparison of two molecular assays for the identification of the SOD1:c.118G>A mutation.

Degenerative myelopathy (DM) is a late-onset, slowly progressive degeneration of spinal cord white matter which is reported primarily in large breed dogs. The missense mutation SOD1:c.118G>A is associated with this pathology in several dog breeds, including the German Shepherd Dog (GSD). The aims of the present study were to develop a tool for the rapid screening of the SOD1 mutation site in dogs and to evaluate the association of the polymorphism with DM in the German Shepherd breed. Two different techniques were compared: a minisequencing test and a real-time pcr allelic discrimination assay. Both approaches resulted effective and efficient. A sample of 47 dogs were examined. Ten subjects presented the symptoms of the illness; for one of them the diagnosis was confirmed by postmortem investigations and it resulted to be an A/A homozygote. In another clinically suspected dog, heterozygote A/G, the histopathological examination of the medulla showed moderate axon and myelin degenerative changes. GSD shows a frequency of the mutant allele equal to 0.17, quite high being a high-risk allele. Because canine DM has a late onset in adulthood and homozygous mutant dogs are likely as fertile as other genotypes, the natural selection is mild and the mutant allele may reach high frequencies. A diagnostic test, easy to implement, may contribute to control the gene diffusion in populations. The SOD1:c.118G>A mutation could be a useful marker for breeding strategies intending to reduce the incidence of DM.

Malignant skin neoplasms in goats in Sicily, Italy: clinical, virological and pathological investigations.

Neoplasms in small ruminants are considered uncommon and their reported incidence is variable. The aims of this investigation were to characterize malignant skin neoplasms in adult goats reared in Sicily, Italy, and to evaluate potential correlations between gross and histopathology features of the tumours and signalment, tumour location and/or viral infections. A total of 75 malignant skin masses were examined. In selected animals with perineal masses (n = 28) virological and serological investigations on tissues and blood were also conducted. According to the histological features, the lesions were classified as 67 squamous cell carcinomas (SCCs) (of which 65 were located in the perineum), six melanomas and two fibrosarcomas. In three cases, neoplasms at the base of the horn were associated with nasal polyps. Among the selected perineal SCCs, papillomaviruses (PVs), caprine herpesvirus 1 and parapoxvirus were not detected on polymerase chain reaction or on serological examination. However, further investigation on a larger sample size is required to evaluate the potential role of PVs in the pathogenesis of skin tumours in goats.

Insect live larvae as a new nutritional model in duck: effects on gut health.

BACKGROUND: This study aimed to evaluate the effects of Hermetia illucens (Black soldier fly-BSF) and Tenebrio molitor (Yellow mealworm-YMW) live larvae as a new nutritional model on duck's gut health, considering gut histomorphometry, mucin composition, cytokines transcription levels, and microbiota. A total of 126, 3-days-old, females Muscovy ducks were randomly allotted to three dietary treatments (6 replicates/treatment, 7 birds/pen): (i) C: basal diet; (ii) BSF: C + BSF live larvae; (iii) YMW: C + YMW live larvae. BSF and YMW live larvae were administered on top of the basal diet, based on the 5% of the expected daily feed intake. The live weight, average daily gain, average daily feed intake and feed conversion ratio were evaluated for the whole experimental period. On day 52, 12 ducks/treatment (2 birds/replicate) were slaughtered and samples of duodenum, jejunum, ileum, spleen, liver, thymus and bursa of Fabricius were collected for histomorphometry. Mucin composition was evaluated in the small intestine through histochemical staining while jejunal MUC-2 and cytokines transcription levels were evaluated by rt-qPCR. Cecal microbiota was also analyzed by means of 16 S rRNA gene sequencing. RESULTS: Birds' growth performance and histomorphometry were not influenced by diet, with a proximo-distal decreasing gradient from duodenum to ileum (p < 0.001), respecting the physiological gut development. Mucin staining intensity and MUC-2 gene expression did not vary among dietary treatments, even though mucin intensity increased from duodenum to ileum, according to normal gut mucus physiology (p < 0.001). Regarding local immune response, IL-6 was higher in YMW group when compared to the other groups (p = 0.009). Insect live larvae did not affect cecal microbiota diversity, but BSF and YMW groups showed a higher presence of Helicobacter, Elusimicrobium, and Succinatimonas and a lower abundance of Coriobacteriaceae and Phascolarctobacterium compared to C birds (p < 0.05). CONCLUSIONS: The use of BSF and YMW live larvae as new nutritional model did not impair gut development and mucin composition of Muscovy ducks, but slightly improved the intestinal immune status and the microbiota composition by enhancing regulatory cytokine IL-6 and by increasing minor Operational Taxonomic Units (OTUs) involved in short-chain fatty acids production.