Use of Vincristine Sulfate in the Treatment of Squamous Cell Carcinoma in a Horse's Lacrimal Gland

Background: Squamous cell carcinoma (SCC) is one of the most common cutaneous neoplasms found in equine breeding. The main cause is still unknown, based on predisposing factors such as excessive exposure to ultraviolet rays, advanced age, depigmented skin and viral or parasitic agents. There is currently a range of possibilities for the treatment of SCC. The choice of the ideal treatment changes according to the location, evolution and size of the tumor, economic costs and availability of material. The present case report aims to report the case of a Campeiro breed, diagnosed with SCC in the left lacrimal gland region and to evaluate the action of vincristine sulfate as intralesional chemotherapy. Case: A 30-year-old stallion of the Campeiro perlino coat it was attended in the sector of Clinic and Surgery of Large Animals, at the Federal University of Santa Catarina - Curitibanos campus. Physical examination revealed an alert and docile temperament, with no considerable physiological changes. In the evaluation of the left eye, it was observed a mass of 2x2 cm rosacea, with granular and ulcerated appearance, it was detected in the portion of the medial corner of the eye, and the lacrimal gland cannot be macroscopically delimited. Also it was detected a moderate mucopurulent secretion. The mass was collected through fine-needle aspiration (FNA) and puncture without aspiration of contents found in the third eyelid region for cytological analysis, and a malignant epithelial cell neoplasia associated with septic suppurative inflammation was observed, suggestive of SCC. After the laboratory results analyzed, 0.25 mL of intralesional vincristine sulfate (Vincizina®) with 20x06 needle was applied in the base of the tumor mass as treatment. After 24 days of the first and only application of the antineoplastic, treatment success was remarkable, with a macroscopic involution of the tumor area up to 80%. Discussion: The anatomical location of SCC covering the lacrimal gland, which is an important mechanism for ocular lubrication and cleaning, has prevented surgical excision from being instituted as an alternative treatment, due to subsequent complications such as corneal dryness and possible ulcer development. Therefore, intralesional chemotherapy became a safe and effective alternative to control and delay tumor mass, preserving the functions of the lacrimal gland. Vincristine is commonly used in veterinary medicine as an antineoplastic treatment of transmissible venereal tumors with inhibitory action on tumor cell division. Previous reports demonstrated success in the use of intravenous vincristine sulfate in the treatment of animals with benign and malignant sarcoids. In the present report satisfactory results were obtained, with only one application of intralesional vincristine sulfate, probably the continuity in the administration of the drug could have propitiated results with more effectiveness. By reason of the surgical excision, treatment of choice, not be feasible in this case, the use of a new form of treatment, less invasive and effective, aims to maintain the function of the affected organ and consequently in the welfare of animal. Every successfully treated animal should be monitored so that, in case of recurrence, it is re-treated as soon as possible

Abordagem laparoscópica na égua como meio auxiliar nas técnicas de reprodução assistida

Laparoscopy is a poorly explored tool for equine assisted reproduction techniques, despite its use in man, zoo and domestic animals. This study had the objective to evaluate the possibility and offer basis for cannulation of the oviduct and its future application in gamete and embryo transfer programs in this species. Seventeen standing laparoscopies were accomplished in ten mares, each ovary approached from its ipsolateral flank. After identification of the uterine tube, the infundibulum was gently tractioned and a catheter advanced in direction of the abdominal ostium of the uterine tube to the ampulla where 250 µL of culture medium (Dulbecco's - PBS) were deposited. The laparoscopic technique associated with assisted reproduction in the mare can be considered pioneer and have the potential of lowering costs with maintenance of receptors by taking advantage of repetitive cycles, which is impossible by means of the conventional approach through laparotomy. This cost is one of the main obstacles for the commercial application of assisted reproduction techniques in horses.A laparoscopia é uma ferramenta pouco explorada nas técnicas assistidas da reprodução em eqüinos, ao contrário do que já ocorre com o homem, animais de zoológicos e domésticos. Este estudo objetivou verificar a possibilidade de oferecer bases para a canulação do oviduto e sua futura aplicação em programas de transferência de gametas e embriões nesta espécie. Foram realizadas 17 laparoscopias em 10 éguas mantidas em estação onde cada ovário foi abordado pelo flanco ipsolateral. Após identificação da tuba uterina, o infundibulum foi gentilmente tracionado e um cateter avançado em direção ao óstio abdominal da tuba uterina para a ampola, onde foram depositados 250 µl de meio de cultivo (Dulbecco's - PBS). A utilização da técnica laparoscópica em reprodução assistida na égua pode ser considerada pioneira e tem o potencial de diminuir os custos de manutenção de receptoras através do aproveitamento de ciclos repetidos, o que não é possível pela abordagem convencional através de laparotomia. Este custo é um dos principais entraves à aplicação comercial das técnicas assistidas da reprodução em eqüinos

Desenvolvimento embrionário de oócitos bovinos mantidos em fluido folicular bovino de folículos de diferentes diâmetros

Bovine oocytes have been maintained in the follicular fluid to be transported and to increase their competence, before maturation. Eight hundred eighty-one (881) oocytes, aspirated from bovine slaughterhouse ovaries, were used to evaluate the effect of holding bovine oocytes in follicular fluid (FF) of bovine follicles of different diameters on the rate of embryo development. The oocytes were randomly distributed in four treatments with seven replicates each: The control group (n=217) was constituted by oocytes matured for 24h in modified TCM-199 with Estrus Mare Serum (EMS), pyruvate and rFSH-h in incubator with 5,00% CO2, 39°C and saturated humidity. In the FFsmall group (3 to 5mm follicles; n=216), the oocytes were held for 6h in follicular fluid at 30°C and matured for 18h in the same conditions of the Control-group. The oocytes of the FFmedium group (5,1-8mm follicles; n=226) and of the FFlarge group (>;8,1mm follicles; n=222) were held in follicular fluid and matured like FFsmall. Fertilization was accomplished during 18h and, after this, the zygotes were cultured for 8 days in SOFaaci medium + 5,00% EMS in incubator at 39°C using plastic bags gasified with 5,00%CO2, 5,00%O2 and 90,00%N2. FFsmall oocytes produced a lower (P;0,05) between the groups. Follicular fluid of medium and large follicles could be used to hold for 6h at 30°C bovine oocytes before their maturation for 18h.Oócitos bovinos têm sido mantidos em fluido folicular como meio para transporte e para aumento de sua competência, antes da maturação. Oitocentos e oitenta e um (881) oócitos foram aspirados de folículos de 2 a 8mm, de ovários de abatedouro, para avaliar o efeito da manutenção de oócitos bovinos em fluido folicular bovino de folículos de diferentes tamanhos sobre o desenvolvimento embrionário. Os oócitos foram distribuídos aleatoriamente em quatro tratamentos, com sete repetições cada. No grupo controle (n=217), os oócitos foram maturados por 24h em TCM-199 com Soro de Égua em Estro (SEE), piruvato e rFSH-h, em estufa a 39°C, com 5,00% de CO2 e umidade saturada. No tratamento FFpequeno (n=216), os oócitos foram mantidos por 6h em fluido folicular de folículos de 3 a 5mm a 30°C e posteriormente maturados por 18h nas mesmas condições do grupo controle. Os oócitos dos tratamentos FFmédio (n=226) e FFgrande (n=222) foram mantidos em fluido folicular de folículos com 5,1 a 8mm e folículos maiores de 8,1mm, respectivamente e, após, maturados por 18h. Após a fecundação por 18h, os zigotos foram cultivados por 8 dias em SOFaaci com 5,00% de SEE, em estufa a 39°C, em bolsas gaseificadas com 5,00%CO2, 5,00%O2 e 90,00%N2. Oócitos do grupo FFpequeno resultaram em menor (P;0,05) entre os grupos. O fluido folicular de folículos médios e grandes pode ser utilizado para o manutenção de oócitos bovinos por 6h a 30°C, antes da maturação por 18h

Produção in vitro de embriões bovinos com soro de égua em diferentes fases do estro

A produção in vitro de embriões (PIV) bovinos é atrativa pelo baixo custo de produção, uso na clonagem de células somáticas e embriões, produção de vacas transgênicas e na pesquisa básica. As proteínas acrescidas aos meios de cultivo parecem ser um dos principais fatores limitantes nas diferentes fases da produção, e são representadas, em primeira instância, por materiais de origem biológica que fornecem aos embriões nutrientes e outros fatores pouco conhecidos. Este trabalho avaliou o uso do soro de égua, obtido em diferentes fase do estro na PIV. Complexos cumulus-oócitos (CCO) obtidos de ovários bovinos em matadouro foram maturados por 22-24h, em estufa (5% de CO2 , umidade saturada e 39oC) em TCM-199 + HEPES + LHb + rFSHh com 10% de soro de égua coletado no primeiro dia de estro (T1), 24-48h antes da ovulação (T2) e 24-48h pós-ovulação (T3), servindo como controle (C) o mesmo meio com soro de vaca em estro. Os CCO foram fecundados em TALP-FERT sob as mesmas condições e cultivados em fluido sintético de oviduto (SOF) + 5% do soro dos T1, T2, T3 ou C, por 8 dias, sob óleo mineral. A taxa de clivagem foi similar entre os tratamentos. A produção de blastocistos no D7 nos tratamentos (T1=21%, T2=21% e T3=20%) foi maior (

Influence of acepromazine on the cardiovascular actions of dobutamine in isoflurane-anesthetized horses Influência da acepromazina sobre os efeitos cardiovasculares da dobutamina em cavalos anestesiados com isofluorano

The influence of acepromazine (ACP) on the effectiveness of dobutamine (DBT) in increasing blood pressure during isoflurane (ISO) anesthesia was evaluated in six horses. On separate occasions, the horses were randomly assigned to receive NaCl 0.9% (Control), ACP 0.025mg kg-1 and ACP 0.05mg kg-1. The experimental treatment was administered prior to induction of anesthesia. Maintenance of anesthesia was performed under conditions of normocapnia with ISO in oxygen. Dobutamine was administered at progressively increasing infusion rates until mean arterial pressure (MAP) reached 70mmHg or until a maximum infusion rate of 5.0µg kg-1 min-1. Compared with baseline, DBT increased heart rate, systolic, diastolic and mean blood pressures in all treatments. However, these variables did not differ among treatments. The target MAP (70mmHg) was not reached in 2/6, 2/5 and 0/6 horses in the Control, ACP0.025 and ACP0.05 treatments, respectively. The mean dose of DBT to achieve target MAP was 3.5±1.8, 3.7±1.6 and 2.7±1.4µg kg-1 min-1 in the Control, ACP0.025 and ACP0.05 treatments, respectively (P>0.05). Under the conditions of this study, premedication with ACP does not interfere with the effectiveness of DBT in increasing blood pressure in horses anesthetized with ISO.A influência da acepromazina (ACP) sobre a capacidade da dobutamina (DBT) em elevar a pressão arterial durante a anestesia com isofluorano (ISO) foi avaliada em seis equinos. Em ocasiões diferentes, os animais receberam aleatoriamente NaCl 0,9% (Controle), ACP 0,025mg kg-1 e ACP 0,05mg kg-1. O tratamento experimental foi administrado previamente à indução da anestesia. A manutenção da anestesia foi realizada em condições de normocapnia com ISO em oxigênio. A administração de DBT foi iniciada em doses progressivamente crescentes até que o valor de pressão arterial média (PAM) atingisse 70mmHg ou até a dose máxima de 5,0µg kg-1 min-1. Comparado ao basal, a administração da DBT resultou em elevação na frequência cardíaca e pressões arteriais sistólica, diastólica e média em todos os tratamentos. Porém, não houve diferença entre os tratamentos nessas variáveis. A PAM alvo (70mmHg) não foi atingida em 2/6, 2/5 e 0/6 animais dos tratamentos Controle, ACP0.025 e ACP0.05, respectivamente. A dose média de DBT para a PAM alvo foi de 3,5±1,8; 3,7±1,6 e 2,7±1,4µg kg-1 min-1 no Controle, ACP0.025 e ACP0.05, respectivamente (P>0,05). Nas condições deste estudo, o pré-tratamento com ACP não interfere na eficácia da DBT em elevar a pressão arterial de cavalos anestesiados com ISO

Approach to Treatment of Multiple Sarcoids in Crioulo Horses

Background: The equine sarcoid corresponds to one third of equine tumors, and may present as single or multiple and in six different shapes. The etiology is still unknown, but it brings with it some predisposing factors. Confirmatory diagnosis is made based on histopathology evaluation. There are several treatments described in the literature, therefore, it varies according to the choice of the veterinarian, the conditions and interests of the owner and the characteristics of the sarcoid tumor. The present research work aims to report the case of a Crioule horse that presented of a multiple sarcoids in different shapes of the body and, testing the efficacy and response to the chosen treatment.Case: The Crioulo gateado coat stallion, aged six years and seven months and weigh 360 kg. In the physical evaluation observed alert behavior, body score (ECC) three (1-5), 36 bpm, 20 mpm. The pelvic right limb (PRL) presented a mobile fibroblastic nodule was found on the proximal dorsal face of the third metatarsal bone (MTIII), pendular and broad-based, ulcerated, with a diameter of approximately 10 cm x 7 cm associated of a dark pink color. Likewise, on the caudal surface of the ear observed verrucous sessile flat lesion with a diameter of 5 cm and, in the right corner of the mouth, a non-ulcerated, pendular, mobile dark red color nodule evidenced with a diameter approximately 3 cm. Similarly, there were multiple ulcerated pendulous dark pink color mobiles nodules in the region of the left and right groin, as well in the pectoral region. For histopathological evaluation, a 0.5 cm specimen tumor of the medial corner of the mouth was collected and observed proliferation of connective tissue and thickening of the epidermis with pseudocarcinomatous projections suggestive of sarcoid lesion tumor. Subsequently, surgical removal tumors associated with cryotherapy and application of 0.25 ml of intralesional vincristine to each region. In addition, an autologous implant tumors specimen of the 0.3 cm approximately was performed after 15 minutes cryopreservation. They were implanted of the subcutaneous neck space in 1 cm incisions and synthesis with simple interrupted pattern using 2-0 nylon. As postoperative PRL care consisted of the utilization Robert Jones bandage for 20 days period, with changes and wound care in an interval of 4 days associated with the daily antisepsis care of the other treated areas. The patient evaluated 10 months after the aforementioned procedures and complete healing tumors lesions.Discussion: The etiology of the sarcoid not well defined, but there are some predispositions such as race, handling of animals and the location of the body. The accompanied animal falls within the aforementioned predispositions observed in this case characterized of the association breed cattle. Probably the sarcoid tumors to be related together of the bovine papillomavirus, and was affected in the most cited regions, such as members, mouth and ears. The sarcoid can appear in six different shapes, several of which can be found in the same animal. Among the treatments mentioned, we opted for the association of surgical removal, associated with cryotherapy and the use of intralesional vincristine, in addition to autologous implants and topical use of Acyclovir. Considering that there have been no recurrences at this moment, the treatment was effective in eliminating the sarcoid tumors and making it a good option for the control lesions of the clinically presented high rates of recurrences

Produção in vitro de embriões bovinos com soro de égua ou de vaca em estro com ou sem a adição de LH/FSH

Mil duzentos e setenta e um oócitos foram divididos em 4 tratamentos com a finalidade de se avaliar a influência da adição de LH e FSH na produção in vitro de embriões bovinos com soro de vaca em estro (SVE) ou soro de éguas obtido no 1masculine dia do estro (SE). Independente do tratamento os oócitos foram maturados com TCM199 + 5,95mg/ml de Hepes, 0,025mg/ml de piruvato de sódio e 2,2mg/ml de bicarbonato de sódio, sendo adicionado 10% de soro de égua (ES), 10% de SVE (VS), 10% de soro de égua + 0,5mg/ml de hormônio luteinizante bovino (LHb) + 0,01UI/ml de hormônio folículo estimulante recombinante humano-rFSHh (EH) e 10% SVE + LHb + rFSHh (VH). Os oócitos assim tratados, foram maturados em estufa com 5% de CO2 a 39masculineC sob umidade saturada por 22-24h. Depois, foram fecundados em TALP-FERT por 18-20h e cultivados por 8 dias em meio SOF + 5% de SE (ES e EH) ou SVE (VS e VH). As taxas de clivagem de 72% obtidas no grupo VH (229/316) e 61% no grupo VS (193/315) foram significativamente menores (p<0,05) que as dos grupos ES (80% - 254/317) e EH (80% - 257/323). A produção de embriões (blastocistos iniciais, blastocistos, blastocistos expandidos e eclodidos) no D7 após a inseminação para os grupos ES (32%), EH (28%) e VH (27%), foi significativamente superior aos 20% obtidos no VS (p<0,05). No D9, verificou-se uma diferença significativa (p<0,05) entre os grupos ES (31%) e EH (29%) quando comparados com o grupo VS (22%), mas não houve diferença quando comparamos com o VH (24%). A taxa de eclosão em D9 foi de 11% (ES), 11% (EH), 10% (VS) e 5% (VH). Não houve diferença entre as médias do número de células dos embriões (Blastocistos expandidos e eclodidos) obtidos no D9. Conclui-se, com estes resultados que, para a obtenção de taxas regulares de blastocistos, não seja necessária a adição de hormônios quando se utilize o soro de égua, e que os hormônios devam ser adicionados quando se utilize soro de vaca