Prenatal Diagnosis of Canine and Feline Twins Using Ultrasound: A Retrospective Study.

Prenatal diagnosis comprises a set of investigations, both instrumental and laboratory-based, which aim to monitor the health of the foetus during pregnancy, from the early stages of embryonic development to the moments preceding delivery. A growing interest is emerging for the preterm ultrasound morphological screening of embryos and foetuses, aimed at assessing the integrity and viability of the conceptus, as well as the early diagnosis of anomalies which can cause complications. This study is a retrospective study of the ultrasonographic findings of twins in the authors' clinical activity from 2016 to 2022. Only seven cases of monochorionic twins were recorded, out of the whole number of evaluations performed on 3120 foetuses, with a prevalence of 0.6% and 0.2% in feline and canine foetuses. All the twins had their own amniotic sac and umbilical cord but presented a single placenta and a single allantoic sac. Unfortunately, the three feline cases were not more recognizable at term. In the four canine cases, three were of opposite sex and then necessarily dizygotic. Twins may have an impact on the success of a pregnancy due to the risk of dystocia, as observed in some of the reported cases. Prenatal ultrasound allows early recognition of twins in dogs and cats.

Comparison between the effects of epidural lidocaine, tramadol, and lidocaine-tramadol on postoperative pain in cats undergoing elective orchiectomy.

BACKGROUND: In veterinary clinical practice, orchiectomy is one of the most common surgical procedures for cats and is performed mainly in young animals. The purpose of this study was to compare three different epidural (EP) analgesic protocols used in cats undergoing orchiectomy in order to determine which protocol resulted in superior outcomes in terms of perioperative analgesia. Twenty-one client-owned male cats were premedicated with a combination of dexmedetomidine (10 µg/kg) and midazolam (0.2 mg/kg) injected intramuscularly. Anesthesia was induced intravenously with propofol. Cats were randomly divided in three treatment groups of seven animals each: Group L received EP lidocaine (2 mg/kg), Group T received EP tramadol (1 mg/kg), and Group LT received EP lidocaine (2 mg/kg) plus tramadol (1 mg/kg). The post-operative pain level was assessed using two different scales: the Glasgow Composite Measure Pain Scale-Feline (CMPS-F) and the Feline Grimace Scale (FGS). Rescue analgesia was administered when the CMPS-F total score was ≥5 or the FGS total score was ≥4. RESULTS: No adverse effects related to tramadol or lidocaine were observed. Based on post-operative pain assessments, significant differences between groups were observed according to both pain scoring systems. In particular, in Group LT, the CMPS-F and FGS scores decreased significantly in the first six hours following castration. CONCLUSIONS: Based on our results, EP lidocaine plus tramadol provided the best post-operative analgesic effects in cats submitted to orchiectomy lasting 6 h and could also be a choice to consider for longer surgical procedures.

Does TLS Exist in Canine Mammary Gland Tumours? Preliminary Results in Simple Carcinomas.

Neoplastic progression is influenced by the expression of tumour antigens that activate an anti-tumour immune response. Human medical studies show that this body defence is carried out in secondary lymphoid organs (SLOs) but also directly in the tumour through organized cellular aggregates that are called tertiary lymphoid structures (TLSs). However, their occurrence has different meanings in different tumour types. For example, the presence of TLSs in breast cancer is associated with the most aggressive subtypes. This paper aimed to study TLSs in canine mammary simple carcinomas. A morphological assessment of the inflammatory infiltrate was performed on H&E sections of fifty cases. Immunohistochemistry was then carried out to typify the inflammatory cells in the tumour microenvironment. Results showed that, sometimes, inflammatory infiltrates were organized in follicles close to high-grade carcinomas, simulating a lymphoid organization, as in breast cancer. Therefore, we can assume that even in canine mammary tumours, TLSs exist and they are entities to consider due to their presence in the most aggressive histotypes or tumours with a high degree of malignancy.

European Regulations on Camel Germplasm Movement within the European Union: A Current Framework Based on Safety.

With the aim of developing livestock breeding, the Delegated Regulation (EU) 2020/686 (hereafter referred to as Reg. 686) has taken steps to define traceability and animal health for the movement of germ material within the European Union (EU), including that of camelid species. Despite the economic importance of the camel market and the efforts of the EU to regulate their movements, there are considerable difficulties in the collection of semen and its freezing, limiting the use of artificial insemination in this species. If, on the one hand, there is little diffusion of the camel breeding and, consequently, limited diffusion of animals and germplasm, there will probably be a significant increase over the years. To avoid the spread of emerging diseases-or even those no longer present in Europe-the entry of genetic material from non-EU countries must be strictly monitored. Camels are rarely clinically compliant, but can transfer even fatal diseases to domestic ungulate farms in the EU. Based on these considerations, we conducted a narrative review of the European regulations on this issue, focusing on aspects related to their application in camels.

Flashing Lights, Dark Shadows, and Future Prospects of the Current European Legislation for a Better Traceability and Animal Health Requirements for Movements of Small Animal Germinal Products.

Recently, there has been an increasing movement of germinal products of dogs (Canis lupus familiaris) and cats (Felis silvestris catus) between the Member States. Therefore, Europe laid down and harmonized rules on the marking of straws and other packages containing germinal products [Commission Delegated Regulation (EU) 2020/686]. Given that germinal products' movement may increase the risk of infectious disease spread, requirements regarding animal health have been revised focusing on control of rabies and echinococcosis, although there are new emerging diseases that may require, even locally, specific requirements. For this reason, veterinarians, operators, and official veterinarians are involved in different phases of the process. Because non-veterinary operators can operate in all phases, they should have a limited role in collecting germinal products, especially for feline species. Veterinarians, instead, should have a main role in the health evaluation of donors, in collecting germ cells with medical techniques and in depositing sperm and embryos with endoscopic or surgical methods. The official veterinarians are the main ones responsible for the application of the rules. This paper aims to provide an overview of the European legislative framework regarding the newly delegated regulation on germinal products in small animals (dogs and cats), highlighting some of the benefits and critical aspects regarding its functioning.

Genotype data not consistent with clonal transmission of sea turtle fibropapillomatosis or goldfish schwannoma.

Recent discoveries of transmissible cancers in multiple bivalve species suggest that direct transmission of cancer cells within species may be more common than previously thought, particularly in aquatic environments. Fibropapillomatosis occurs with high prevalence in green sea turtles ( Chelonia mydas) and the geographic range of disease has increased since fibropapillomatosis was first reported in this species. Widespread incidence of schwannomas, benign tumours of Schwann cell origin, reported in aquarium-bred goldfish (Carassius auratus), suggest an infectious aetiology. We investigated the hypothesis that cancers in these species arise by clonal transmission of cancer cells. Through analysis of polymorphic microsatellite alleles, we demonstrate concordance of host and tumour genotypes in diseased animals. These results imply that the tumours examined arose from independent oncogenic transformation of host tissue and were not clonally transmitted. Further, failure to experimentally transmit goldfish schwannoma via water exposure or inoculation suggest that this disease is unlikely to have an infectious aetiology.

Bovine besnoitiosis in a cattle herd in Sicily: an isolated outbreak or the acknowledgment of an endemicity?

Bovine besnoitiosis is a debilitating infectious disease caused by Besnoitia besnoiti (Apicomplexa; Sarcocystidae). The disease is mainly characterized by cutaneous and systemic signs, infertility in bulls, and abortion in cows. The current study describes an autochthonous outbreak of bovine besnoitiosis in Sicily, Southern Italy, being the first report of B. besnoiti infection in the island so far. In a cattle farm located in Syracuse province, a 4-year-old Belgian blue bull born in Sicily displayed typical clinical signs of the sub-acute/chronic disease phase with thickening of the skin of the scrotum and testicular degeneration. Histological examination of scrotal biopsies revealed the presence of several tissue cysts of B. besnoiti. The serological analysis of the herd using a commercial ELISA revealed a high seroprevalence (45 out of 55; 82%) of antibodies against B. besnoiti. Few seropositive animals (5 out of 45; 11%) showed clinical signs, cysts in vestibulum vaginae (1 out of 31; 3.2%), and testicular degeneration (4 out of 14; 28.6%) assessed by ultrasonographic investigations. The paucity of clinical signs associated with the high seroprevalence in the farm led to hypothesize that bovine besnoitiosis is endemic in the area though further studies are needed. Local practitioners should be more aware of the disease to facilitate the early detection of cases, prevent the spread of infection, and avoid economic losses and animal health problem.

Constant Rate Infusion of Lidocaine, Tumescent Anesthesia and Their Combination in Dogs Undergoing Unilateral Mastectomy.

Tumescent anesthesia (TUM) is a technique that was initially used to perform liposuction under local anesthesia, which consists of the injection of such large volumes of local anesthetic until to produce swelling and firmness (tumescence) of the surgical area. The aim of this study was to compare the intraoperative analgesic efficacy of lidocaine (LID) constant rate infusion (CRI), of TUM, or their combination (LID/TUM) and the postoperative pain and analgesic requirement in dogs undergoing unilateral mastectomy. Twenty-four dogs were premedicated with dexmedetomidine (3 µg/kg) and methadone (0.2 mg/kg) intravenously (IV). After induction with propofol IV to effect, dogs were randomly allocated to receive a loading dose of lidocaine (2 mg/kg) followed by a CRI of 100 µg/kg/min (Group LID) in addition to an equivalent volume of lactated Ringer's solution instead of local TUM; a loading dose of lactated Ringer's solution followed by a CRI of Ringer's solution in addition to TUM (Group TUM); a loading dose of lidocaine (2 mg/kg) followed by a CRI of 100 µg/kg/min in addition to TUM (Group LID/TUM). Anesthesia was maintained with isoflurane in oxygen. Postoperative pain scores were assessed once the dogs had fully recovered from the sedative effects, and following 15, 30, 45 and 60 min. The results of the current study allow us to assert that all three treatments provided satisfactory intraoperative antinociceptive effects but administration of LID/TUM induced greater inhibition on sympathetic stimulating effect up to 60 min from recovery, thus, providing better early postoperative pain relief in dogs undergoing mastectomy.

Imaging of Canine Neoplastic Reproductive Disorders.

Diagnostic imaging plays an essential role in the diagnosis and management of reproductive neoplasia in dogs and cats. The initial diagnosis, staging, and planning of surgical and radiation treatment and the response to therapy all involve imaging to varying degrees. Routine radiographs, ultrasound, nuclear medicine, and cross-sectional imaging in the form of computed tomography (CT) and magnetic resonance imaging (MRI) are routinely used in canine reproductive disorders. The choice of imaging modality depends on many factors, including the level of referral and the pathological information required. The biological behaviour of the tumour also guides the choice of imaging in cancer staging, and imaging may play an important role in guiding serial tumour biopsy during the course of therapy. The sophistication of imaging modalities is increasing exponentially. Each modality has advantages and disadvantages in terms of cost, availability, sensitivity, specificity, and qualities of anatomic versus functional imaging.

Immune Cells and Immunoglobulin Expression in the Mammary Gland Tumors of Dog.

Inflammatory cells have a role in tumor progression and have prognostic and therapeutic potential. The immunohistochemical expression for Mast Cell Tryptase, Macrophage Marker, CD79a, IgA, IgM and IgG on 43 cases of canine mammary gland lesions was analyzed. In hyperplasia, a few B cells (BCs) and Tumor-Associated Macrophages (TAMs) were observed, while the number of Tumor-Associated Mast Cells (TAMCs) was the highest. In the peritumoral stroma of malignant lesions, low number of TAMCs and a high number of TAMAs and BCs were present. Immune cells of each type were always lower in the intratumoral than peritumoral stroma. Positivity to CD79a was also detected in the epithelial cells of simple and micropapillay carcinomas. Immunoglobulin reactivity was mainly located in the epithelial cells where an intense positivity to IgA and IgG and a weak positivity for IgM were detectable. On the basis of our preliminary results and literature data, we suggest that such cells and molecules could be directly involved in the biology of canine mammary gland tumors. In breast cancer, stromal inflammatory cells and cancer derived immunoglobulins have been correlated with the progression, malignancy and poor prognosis of the tumor. The results herein reported show that the dog's mammary gland epithelium also expresses immunoglobulins, and they mostly show a direct relationship with the infiltration of macrophages. In addition, this study shows that the infiltration of mast cells, B-cells and macrophages varies depending on the degree of malignancy of neoplasia.

Pseudo-Placentational Endometrial Hyperplasia in the Bitch: Case Series.

Canine pseudo-placentational endometrial hyperplasia differs from the classical form of cystic endometrial hyperplasia for the well-organized tissue architecture resembling the canine placenta. After the discovery, it has been inconstantly reported. The present work reports the clinicopathological details of six spontaneous cases retrieved retrospectively from a large database. The lesion was found in young non-pregnant female dogs (median 2.0 years) at the end of dioestrus. It could be imaged by ultrasound and was always grossly detectable as single or multiple uterine enlargements of 2-3 cm in diameter with a villous whitish tissue growing on the mucosa and occluding the lumen. Histology confirmed the tissue architecture of the canine placenta with a basal glandular layer, a connective band, a spongy layer and a tortuous and compact labyrinth, often poorly recognizable. The pseudo-placentational hyperplasia is a non-inflammatory proliferative lesion although numerous mast cells inhabit the connective band, and a superimposed inflammatory infiltrate was seen in a case. Canine pseudo-placentational endometrial hyperplasia has very peculiar features, and it is a model for canine placentation and may help to better understand the cystic endometrial hyperplasia/pyometra complex.

Welfare assessment in intensive and semi-intensive dairy cattle management system in Sicily.

The present study aimed to compare the welfare of dairy cows kept in two traditional husbandry systems (semi-intensive and intensive farming) in south-eastern Sicily. A total of 18 dairy farms (nine semi-intensive and nine intensive) were evaluated with a multicriteria system adapted for Sicilian conditions and obtained simplifying the model of the European Food Safety Authority (EFSA). Values of welfare measures, collected by inspections of the farms (general well-being indicators, ventilation system, resting areas [cubicles or bedding], flooring, milking parlours and waiting area, manger and watering equipment), and those of health categories (cases of abortions, hypocalcemia, displacement of abomasum, acidosis/ketosis, enteritis, hoof problems, and mastitis) obtained through the farm records, were compared using Mann-Whitney and Chi-squared tests, respectively. Data showed significant differences (p ≤ .05) about the variables related to welfare categories such as housing ventilation system, resting area, manger, and water equipment that were better in the semi-intensive system than the intensive system. No significant differences were observed about the variables related to health indicators. The results demonstrated that in Sicily the semi-intensive farm is better than the intensive to satisfy the conditions of animal welfare.

Use of GnRH Agonist in Dogs Affected with Leishmaniosis.

Sex-associated hormones such as testosterone have been demonstrated to modulate immune responses, which can result in different disease outcomes. The present study was aimed at evaluating the efficacy of a gonadotropin-releasing hormone GnRH agonist implant as deslorelin acetate in association with meglumine antimoniate plus allopurinol in dogs with canine leishmaniosis (CanL). Twenty-two dogs with CanL confirmed by clinical findings and laboratory tests were included in the study. Dogs were randomized into two groups. A control group (CTR, n = 12) was treated with meglumine antimoniate 50 mg/kg SC q 12 h for 28 days plus allopurinol at 10 mg/kg PO q 12 h for the whole study period (six months). An experimental group was treated with allopurinol and meglumine antimoniate, plus an implant of 4.7 mg deslorelin acetate (DES, n = 10). The animals were observed for three months, during which clinical evaluation, indirect fluorescent antibody test (IFAT) titre and testosterone assay were performed on time at day (D)0, 90 and 180. A significantly lower clinical score was recorded in DES than in CTR (p < 0.01) at D90 and D180 (p < 0.01). After 180 days of treatment (D180), a significant reduction of mean levels of IFAT was observed in the DES group (p = 0.03). A highly significant reduction of testosterone (p = 0.01) was observed in the DES group during the study. No statistical correlation between clinical scores, IFAT titres and testosterone within two groups was observed. Data suggested that the agonist of GnRH may be useful in the treatment of CanL.

Conservative treatments for feline fibroadenomatous changes of the mammary gland.

BACKGROUND: Fibroadenomatous changes (FAC) of the mammary gland in cats represents a benign, progesterone-associated fibroglandular proliferation of one or more mammary glands that may occur in intact female cats at the time of puberty, during pregnancy or pseudopregnancy, or in female and male cats of any age under progestin treatment. Nowadays, the elective treatment of FAC is based on the progesterone antagonist aglepristone. This study aimed to report the treatment of FAC with a combination of drugs designed to preserve mammary gland integrity, even in pregnant cats. CASE DESCRIPTION: Eight sexually intact female cats with FAC showed mammary glands that were symmetrically enlarged and inflamed in all cases, and the skin was ulcerated in six cats. Four cats were on days 25-32 of pregnancy at presentation. Non-pregnant cats were treated with aglepristone and with a dietary supplement containing maltodextrin and bromelain. The mammary glands were massaged daily with an Aloe vera emollient gel. If the gland was inflamed or ulcerated, broad-spectrum antimicrobial, and anti-inflammatory treatments were given, and the ulcers were treated topically with a hypericum and neem-based cream. Two of the four pregnant cats were treated with the same therapeutic schedule plus cloprostenol to facilitate uterine emptying. Two pregnant cats underwent the same protocol except for aglepristone, which was not administered to safeguard the litter and see if the adjuvant therapy worked independently. At term, they delivered four and three kittens, respectively, that were normally nursed and weaned after 40 days. In all the studied cases, the mammary gland reduced in size 2-3 weeks after the start of the treatment and completely remitted after 4-5 weeks. CONCLUSION: This case series encourages adjuvant therapy in the course of FAC to preserve mammary gland integrity and functionality. Exploiting the natural decline of progesterone at the end of pregnancy (or pseudopregnancy), the therapy may also be used without aglepristone, when its use has other limitations.

Identification of trypanosomatids and blood feeding preferences of phlebotomine sand fly species common in Sicily, Southern Italy.

In this study, the presence of Leishmania DNA and blood feeding sources in phlebotomine sand fly species commonly present in Sicily were investigated. A total of 1,866 female sand flies including 176 blood fed specimens were sampled over two seasons in five selected sites in Sicily (southern Italy). Sergentomyia minuta (n = 1,264) and Phlebotomus perniciousus (n = 594) were the most abundant species at all the sites, while three other species from the genus Phlebotomus (i.e., P. sergenti n = 4, P. perfiliewi n = 3 and P. neglectus n = 1) were only sporadically captured. Twenty-eight out of the 1,866 (1.5%) sand flies tested positive for Leishmania spp. Leishmania tarentolae DNA was identified in 26 specimens of S. minuta, while the DNA of Leishmania donovani complex was detected in a single specimen each of S. minuta and P. perniciosus. Interestingly, seven S. minuta specimens (0.4%) tested positive for reptilian Trypanosoma sp. Blood sources were successfully identified in 108 out of 176 blood fed females. Twenty-seven out of 82 blood sources identified in fed females of P. perniciosus were represented by blood of wild rabbit, S. minuta mainly fed on humans (16/25), while the sole P. sergenti fed specimen took a blood meal on rat. Other vertebrate hosts including horse, goat, pig, dog, chicken, cow, cat and donkey were recognized as blood sources for P. perniciosus and S. minuta, and, surprisingly, no reptilian blood was identified in blood-fed S. minuta specimens. Results of this study agree with the well-known role of P. perniciosus as vector of L. infantum in the western Mediterranean; also, vector feeding preferences herein described support the hypothesis on the involvement of lagomorphs as sylvatic reservoirs of Leishmania. The detection of L. donovani complex in S. minuta, together with the anthropophilic feeding-behaviour herein observed, warrants further research to clarify the capacity of this species in the transmission of pathogens to humans and other animals.

Somatic evolution and global expansion of an ancient transmissible cancer lineage.

The canine transmissible venereal tumor (CTVT) is a cancer lineage that arose several millennia ago and survives by "metastasizing" between hosts through cell transfer. The somatic mutations in this cancer record its phylogeography and evolutionary history. We constructed a time-resolved phylogeny from 546 CTVT exomes and describe the lineage's worldwide expansion. Examining variation in mutational exposure, we identify a highly context-specific mutational process that operated early in the cancer's evolution but subsequently vanished, correlate ultraviolet-light mutagenesis with tumor latitude, and describe tumors with heritable hyperactivity of an endogenous mutational process. CTVT displays little evidence of ongoing positive selection, and negative selection is detectable only in essential genes. We illustrate how long-lived clonal organisms capture changing mutagenic environments, and reveal that neutral genetic drift is the dominant feature of long-term cancer evolution.

Molecular Signatures of Regression of the Canine Transmissible Venereal Tumor.

The canine transmissible venereal tumor (CTVT) is a clonally transmissible cancer that regresses spontaneously or after treatment with vincristine, but we know little about the regression mechanisms. We performed global transcriptional, methylation, and functional pathway analyses on serial biopsies of vincristine-treated CTVTs and found that regression occurs in sequential steps; activation of the innate immune system and host epithelial tissue remodeling followed by immune infiltration of the tumor, arrest in the cell cycle, and repair of tissue damage. We identified CCL5 as a possible driver of CTVT regression. Changes in gene expression are associated with methylation changes at specific intragenic sites. Our results underscore the critical role of host innate immunity in triggering cancer regression.

Mitochondrial genetic diversity, selection and recombination in a canine transmissible cancer.

Canine transmissible venereal tumour (CTVT) is a clonally transmissible cancer that originated approximately 11,000 years ago and affects dogs worldwide. Despite the clonal origin of the CTVT nuclear genome, CTVT mitochondrial genomes (mtDNAs) have been acquired by periodic capture from transient hosts. We sequenced 449 complete mtDNAs from a global population of CTVTs, and show that mtDNA horizontal transfer has occurred at least five times, delineating five tumour clades whose distributions track two millennia of dog global migration. Negative selection has operated to prevent accumulation of deleterious mutations in captured mtDNA, and recombination has caused occasional mtDNA re-assortment. These findings implicate functional mtDNA as a driver of CTVT global metastatic spread, further highlighting the important role of mtDNA in cancer evolution.

Transmissible [corrected] dog cancer genome reveals the origin and history of an ancient cell lineage.

Canine transmissible venereal tumor (CTVT) is the oldest known somatic cell lineage. It is a transmissible cancer that propagates naturally in dogs. We sequenced the genomes of two CTVT tumors and found that CTVT has acquired 1.9 million somatic substitution mutations and bears evidence of exposure to ultraviolet light. CTVT is remarkably stable and lacks subclonal heterogeneity despite thousands of rearrangements, copy-number changes, and retrotransposon insertions. More than 10,000 genes carry nonsynonymous variants, and 646 genes have been lost. CTVT first arose in a dog with low genomic heterozygosity that may have lived about 11,000 years ago. The cancer spawned by this individual dispersed across continents about 500 years ago. Our results provide a genetic identikit of an ancient dog and demonstrate the robustness of mammalian somatic cells to survive for millennia despite a massive mutation burden.

Sertoli cell tumors associated with feminizing syndrome and spermatic cord torsion in two cryptorchid dogs.

The association of cryptorchidism, functional Sertoli cell tumors, and spermatic cord torsion has been rarely reported in the literature. Two dogs were admitted for bilateral skin alopecia and weight loss. Both animals were cryptorchid and displayed a pendulous preputial sheath, prostate hypertrophy, and increased levels of circulating oestrogen. Transabdominal palpation and ultrasonography revealed the presence of neoplastic retained gonads. During surgery, spermatic cord torsion was also detected in the enlarged neoplastic testes of both dogs. Histologic examination confirmed the presence of Sertoli cell tumors that were primarily responsible for the feminizing syndrome. Complete remission of all symptoms occurred within 3 months after orchiectomy.