Avian coccidiosis caused by Eimeria spp. is a relevant disease for poultry usually prevented by synthetic coccidiostats. Citrus aurantium L. var. dulcis peel essential oil (CEO), and Yucca schidigera's saponins (Ys) were tested as valid natural alternatives to coccidiostats drugs. Their anticoccidial effects, gut health and growth performance of broiler chickens were investigated with discovery-driven research by a field trial during the broiler chicken production cycle in a rural free-range farm with a history of coccidiosis infection. One hundred forty 1-d-old female broiler chickens were assigned to 4 experimental treatment groups (35 broiler chicks each), fed ad libitum with a coccidiostats-free diet, during 9 wk of the trial (63 d). The treatment groups included: control (C); CEO (1 ml/L); Ys pure extract (0.05 ml/L); and CEO + Ys (CEO 1 ml/L + Ys 0.05 ml/L), administered by drinking water. Fecal coccidia oocyst count (OPG), and growth performance were evaluated weekly. On wk 9 all broiler chickens were slaughtered, and serum biochemical indices and intestinal lesions were histologically evaluated and scored. Results showed that from the 6th to 9th wk, the treated groups had lower OPG than the C group (P < 0.05), with the lower value observed in the CEO and Ys groups. More severe coccidiosis intestinal lesions attributable to E. maxima were detected in the C group. All chickens showed a variable degree of intestinal lymphoplasmacytic and heterophilic inflammation without differences in the histological and intestinal lesion scores. At the end of the trial, the body weight (BW) was greater in the treated groups compared to the C group (P < 0.05). However, no differences in FI, WDI, FCR, mortality, and organ weights were evidenced. Albeit within the physiological range, serum AST, cholesterol, total proteins, albumin, and P were higher in the Ys group (P < 0.05), compared to the C group, which was also observed in the CEO + Ys group for the cholesterol, total protein and albumin content. The CEO and Ys biologically active metabolites, even if taken in moderate concentrations, can help to reduce the OPG and gut damage during the coccidia infection, improving the broiler chicken's growth performance.

Canine splenic hemangiosarcoma has a high metastatic rate and short survival time. Currently, the main prognostic parameters are tumor stage and therapy, while data on histologic parameters, such as grade and Ki-67 expression, are scarce. The aims of this study were to compare two methods of assessment of Ki-67, verify their prognostic impact, and define a threshold value based on survival. Thirty-one cases of histologically diagnosed canine splenic hemangiosarcoma, which were treated with splenectomy and had full staging and follow-up information, were collected. Three were stage I, 17 stage II, and 11 stage III. The mean mitotic count (MC) was 23.9 (standard deviation [SD]: 22.1) and the median was 15 (range, 1-93). Immunohistochemistry for Ki-67 was performed, the Ki-67 labeling index (Ki-67LI) was assessed as a percentage of positive neoplastic nuclei per ≥500 cell, and the Ki-67 count (KI-67C) was defined as the average number of positive nuclei using a 1 cm2 optical grid performed in 5, 40× fields. The mean Ki-67LI and Ki-67C were 56.4% (SD: 38.7) and 27.2 (SD: 12.9) and medians were 51% (range, 8.2-55.2) and 26 (range, 5.5-148), respectively. Using a cut-off of 56% and 9, respectively, Kaplan-Meier survival curves showed an association of overall survival with Ki-67LI and MC. In addition to clinical stage, Ki-67LI maintained its prognostic value on multivariate analysis, supporting the role of Ki-67LI as an independent prognostic parameter. Based on these results, we propose a diagnostically applicable cut-off value of 56% for Ki-67LI as a prognostic parameter for canine splenic hemangiosarcoma.

Soft tissue sarcomas (STSs) are conventionally viewed as poorly immunogenic tumors; however, some human STSs have recently been reported to elicit an immune response, thus representing potential candidates for immunotherapy. Data regarding immune cell infiltrates in canine STSs are limited and reported without tumor-type stratification. The aim of this study was to retrospectively assess tumor-infiltrating lymphocytes (TILs) in canine STSs of 5 different histotypes. Eighty-seven canine STSs were collected: 22 perivascular wall tumors (PWTs), 19 liposarcomas, 17 fibrosarcomas, 16 myxosarcomas, and 13 leiomyosarcomas. The tumors were graded and immunolabeled for CD3, CD20, and FoxP3, and slides were scanned. T-cell, B-cell, Treg, and total TIL densities were quantified with QuPath software and expressed as cells/mm2. The B/T-cells ratio and Treg/T-cell proportions were calculated. Total TIL densities were higher in PWTs and myxosarcomas (median = 225 and 303, respectively). PWTs had higher T-cell density but lower Treg proportion (median = 152 and 7.6% respectively). Myxosarcomas had higher Treg densities and B/T-cell ratios (median = 24.4 and 1.57, respectively). No association with grade was found among STSs as a group. In myxosarcomas, higher grade was significantly associated with higher total TILs, and CD20+ and FoxP3+ cell densities ( p < .05). The results suggest that PWTs and myxosarcomas may represent the most immunogenic STS types. Myxosarcomas elicit a B-cell and Treg-rich immune response; PWTs stimulate a T-cell-rich and Treg-poor reaction. The immune system response may contribute to the more aggressive behavior of myxosarcomas and the more indolent course of PWTs.

Avian coccidiosis caused by Eimeria spp. is a relevant disease for poultry usually prevented by synthetic coccidiostats. Citrus aurantium L. var. dulcis peel essential oil (Ca-EOs) and Yucca schidigera’s saponins (Ys) are tested as a valid natural alternatives to coccidiostats. A field study was conducted during the broiler production cycle to evaluate their effects on broiler reared in a rural free-range farm exposed to environmental coccidia infection. One-day-old female broiler chicks (n=140) were divided into 4 experimental groups (n=35 each), fed ad libitum with a coccidiostats-free diet, during 9 weeks of trial, from 1 (63 days). The treatment groups included: control (C); Ca-EOs (1 ml/l); Ys pure extract (0.05 ml/l); Ca-EOs+Ys (1.05 ml/l), administered by drinking water. Fecal coccidia oocysts count (OPG), and growth performance were evaluated weekly. At week 9 all animals were slaughtered, and serum biochemical parameters and intestinal lesions were histologically evaluated and scored. The orange EOs from peel waste and yucca saponins showed a coccidiostat-like effect maintaining the coccidiosis in the subclinical state. Results showed that from the 4th to 9th week, treated groups had significantly lower OPG than C (P < 0.05), with the lower value observed in Ca-EOs. All chickens showed a variable degree of intestinal lymphoplasmacytic and heterophilic inflammation without differences in the histological and intestinal lesion scores (P > 0.05). The body weight (BW) was greater in the treated groups compared to the C group (P < 0.05). However, no differences in FI, WDI, FCR, and mortality were evidenced. Albeit within the physiological range, serum AST, cholesterol, proteins, albumin, Ca, and P were higher in Ys and Ca-EOs+Ys groups (P < 0.05). The Ca-EOs and yucca saponins’ biologically active metabolites, even if taken in moderate concentrations, can help to reduce the coccidia life cycle and gut damage during the environmental coccidia infection, improving the chicken's performance.

Sarcoids are among the most common tumors diagnosed in equids; their association with bovine papillomaviruses (BPV) infection has been widely reported, but the mechanism of carcinogenesis has not been fully elucidated. To verify whether BPV infection causes dysregulation of the pRb-Cyclin D1-p16CDKN2A-p53 pathway as reported for human papillomavirus (HPV), the study employed immunohistochemistry to test 55 equine sarcoid biopsies for the expression of pRb, Cyclin D1, and p53 cell cycle regulatory proteins and to evaluate the proliferative rate through Ki67. High Cyclin D1 and pRb expression were observed in 51% and 80% of cases, respectively, while low expression was observed in 49% and 20% of cases, respectively. Significantly higher Ki67 proliferation indexes were observed in fibroblastic, nodular, and mixed sarcoids compared to the occult and verrucous. High proliferation was significantly associated with high Cyclin D1 expression. In contrast with previous studies, p53 positivity was not observed in the cases examined in this study. Moreover, follow-up analysis revealed that fibroblastic, mixed sarcoids were associated with significantly higher local recurrence rates while the verrucous subtype was associated with higher rates of new sarcoid development at distant sites.