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Embryonic mortality in cattle is high, reaching 10-40 % in vivo and 60-70 % in vitro. Death of embryos involves reduced expression of genes related to embryonic viability, inhibition of DNA repair and increased DNA damage. In follicular granulosa cells, FGF18 from the theca layer increases apoptosis and DNA damage, so we hypothesized that FGF18 may also affect the oocyte and contribute to early embryonic death. The aims of this study were to identify the effects of FGF18 on cumulus expansion, oocyte maturation and embryo development from cleavage to blastocyst stage using a conventional bovine in vitro embryo production system using ovaries of abattoir origin. Addition of FGF18 during in-vitro maturation did not affect FSH-induced cumulus expansion or rates of nuclear maturation. When FGF18 was present in the culture system, rates of cleavage were not affected however, blastocyst and expanded blastocyst development was substantially inhibited (P < 0.05), indicating a delay of blastulation. The number of phosphorylated histone H2AFX foci per nucleus, a marker of DNA damage, was higher in cleavage-stage embryos cultured with FGF18 than in those from control group (P < 0.05). Furthermore, FGF18 decreased accumulation of PTGS2 and IFNT2 mRNA in blastocysts. In conclusion, these novel findings suggest that FGF18 plays a role in the regulation of embryonic death during the early stages of development by impairing DNA double-strand break repair and expression of genes associated with embryo viability and maternal recognition of pregnancy during the progression from oocyte to expanded blastocysts.

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Cumulus cells from cumulus-oocyte complexes (COC) matured in vitro in serum-free medium show high incidence of apoptosis and DNA double-strand breaks (DSB). This study aimed to characterize the transcript expression profile of selected genes involved in DNA repair mechanisms in bovine cumulus cells cultured with bovine serum albumin (BSA) or fetal calf serum (FCS). Briefly, bovine cumulus-oocyte complexes were in vitro matured with either, 0.4% BSA or 10% FCS for 3, 6, 12 or 24 h. The total RNA of cumulus cells was used for real-time PCR analysis. Transcript abundance of XRCC6, XRCC5, DNAPK, GAAD45B, TP53BP1, RAD50, RAD52, ATM and BRCA2 target genes changed as the IVM proceeded (P < 0.05). However, an interaction between protein source (FCS or BSA) and time was not detected (P ≥ 0.05). Cumulus cells from COCs matured with BSA presented higher mRNA expression of two genes compared to FCS group: TP53BP1 at 6 h and BRCA1 at 3, 6, 12 and 24 h (P < 0.05). In summary, our results showed for the first time the expression profile of the key genes involved in DSB repair mechanisms in cumulus cells obtained from bovine COCs matured with FCS or BSA. The higher mRNA expression of BRCA1 and TP53BP1 and lower mRNA expression of TNFAIP6 suggests an increase in apoptosis rate and DNA damage in cumulus cells cultured in BSA-supplemented medium and may explain, at least to some extent, the reduced developmental potential of bovine oocytes matured in serum-free medium.

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Background: Lacrimal gland adenocarcinoma is an extremely rare malignant neoplasm, with few descriptions in the literature. Therefore, its etiology and treatment are not well understood. The present study aims to report the case of an equine histologically diagnosed with lacrimal gland adenocarcinoma in the lower eyelid region and third eyelid treated by surgical excision and intralesional chemotherapy. Case: A 17-year-old male mixed-breed equine weighing 300 kg was treated in the large animal clinic and surgery sector of the Federal University of Santa Catarina in the city of Curitibanos, Santa Catarina, Brazil. The equine revealed an alert and docile temperament without considerable physiological changes in the physical examination. In the evaluation of the right eye, a tumor mass was observed in the region of the lacrimal gland adhered to the third eyelid and lower eyelid, with a light red ulcerated appearance approximately 7 cm in diameter. Therefore, the surgical excision of the adhered tumor mass was performed, with the subsequent intralesional application of 2 mL of Vincristine Sulfate. In the histopathological evaluation following the surgical excision of the tumor mass, yellowish-white fragments of irregular nodules were observed measuring from 3.5×2.0×1.7 cm to 2.0×0.5×0.3 cm, in addition to microscopy focus clusters of weakly basophilic neoplastic glandular cells, some with a randomly arranged lacy aspect, forming disorganized acinar structures and others showed marking islands of the cells organized in a palisade shape by vascular delicate stroma. The neoplastic cells presented anisocytosis, anisokaryosis, prominent nucleoli sometimes binucleated and with basophilic intracytoplasmic secretory material, delimited by scarce connective tissue. In some areas, there were small foci of infiltrated lymphocytes and plasmocytes and areas of necrosis. The mass was surrounded by connective tissue, where red blood vessels were found outside the vessels (hemorrhage), and mitoses were observed 2 per field at high magnification (40x). After the surgical procedure, it was not possible to follow up and reassess of the patient, so there is no clarification on the possibility of tumor recurrence. Discussion: The case reported is uncommon, being only the second report of adenocarcinoma in the equine lacrimal gland. Therefore, there are few descriptions in the literature about its defined etiology and the best treatment method, although, surgical excision is the method of choice because it is a tumor of high recurrence and invasiveness. In this case, we opted for the exeresis of the tumor mass and later application of chemotherapy, a treatment that was relatively effective, to avoid recurrence and the failure of the technique. In these cases, monitoring the animal after the procedure is recommended, evaluating whether there was tumor recurrence, although this was not possible in the present possible. The third eyelid, lacrimal, and zygomatic salivary glands may be sources of intraorbital neoplasms. These neoplastic processes may have similar histological and behavioral characteristics, and their differentiation is, therefore, problematic. These structures may be differentiated based on the anatomical location, however, clinical appearance and symptoms are identical in most cases. Adenocarcinoma in the equine lacrimal gland is a neoplasm considered rare, so it is important to emphasize an accurate diagnosis through histopathological analyses to differentiate it from other frequent orbital neoplasms in horses, allowing more information about this tumor and establishing different treatment methods.

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Background: The equine sarcoid corresponds to one third of equine tumors, and may present as single or multiple and insix different shapes. The etiology is still unknown, but it brings with it some predisposing factors. Confirmatory diagnosisis made based on histopathology evaluation. There are several treatments described in the literature, therefore, it variesaccording to the choice of the veterinarian, the conditions and interests of the owner and the characteristics of the sarcoidtumor. The present case report describe of a Crioulo horse that presented of a multiple sarcoids in different shapes of thebody and, testing the efficacy and response to the chosen treatment.Case: The Crioulo gateado coat stallion, aged six years and seven months, weigh 360 kg. In the physical evaluation observed alert behavior, body score (ECC) three (1-5), 36 bpm and 20 mpm. The pelvic right limb (PRL) presented a mobilefibroblastic 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 ofthe 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 multipleulcerated pendulous dark pink color mobiles nodules in the region of the left and right groin, as well in the pectoral region.Histopathological evaluation of the 0,5 cm specimen tumor observed a proliferation of connective tissue and thickening ofthe epidermis with pseudocarcinomatous projections suggestive of sarcoid lesion tumor...(AU)

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Background: The equine sarcoid corresponds to one third of equine tumors, and may present as single or multiple and insix different shapes. The etiology is still unknown, but it brings with it some predisposing factors. Confirmatory diagnosisis made based on histopathology evaluation. There are several treatments described in the literature, therefore, it variesaccording to the choice of the veterinarian, the conditions and interests of the owner and the characteristics of the sarcoidtumor. The present case report describe of a Crioulo horse that presented of a multiple sarcoids in different shapes of thebody and, testing the efficacy and response to the chosen treatment.Case: The Crioulo gateado coat stallion, aged six years and seven months, weigh 360 kg. In the physical evaluation observed alert behavior, body score (ECC) three (1-5), 36 bpm and 20 mpm. The pelvic right limb (PRL) presented a mobilefibroblastic 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 ofthe 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 multipleulcerated pendulous dark pink color mobiles nodules in the region of the left and right groin, as well in the pectoral region.Histopathological evaluation of the 0,5 cm specimen tumor observed a proliferation of connective tissue and thickening ofthe epidermis with pseudocarcinomatous projections suggestive of sarcoid lesion tumor...

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Hemoglobin is a protein present in erythrocytes, used in the transport of respiratory gases. This protein can be measured through laboratory tests, thus it is an important tool in health assessment of domestic animals. The hemoglobin test protocol by cyano-met hemoglobin method describes that its measurement can be made during a period of up to seven days, but the laboratorial routine shows a reduction in the values before this period. The objective of this study was to determine what the actual maximum period to measure hemoglobin levels in cattle is without changes. Nineteen healthy bovine animals were used, males and females, who underwent venous blood collection via jugular puncture. Samples were analyzed by manual colorimetric. Analyses were made every 24 hours during a fourteen days-interval. During this interval the samples were kept under refrigeration. Hemoglobin levels varied from 8.546 to 22.456 g/dL. Compared to baseline, there were no significant changes in hemoglobin concentration during the investigated period of time, indicating that its measurement can be performed after 14 days of storage.(AU)

A hemoglobina é uma proteína presente nos eritrócitos, que realiza o transporte dos gases respiratórios. Exames laboratoriais mensuram esta proteína, sendo uma importante variável para a avaliação de saúde em animais domésticos. O protocolo do teste da determinação da concentração de hemoglobina pelo método ciano-metemoglobina descreve que sua mensuração pode ser realizada em um prazo de até sete dias, porém a rotina laboratorial mostra redução nos valores antes do período proposto. O objetivo deste trabalho foi avaliar qual o real prazo máximo para mensurar as concentrações de hemoglobina em bovinos sem que haja alterações em seus valores. Foram utilizados 19 bovinos sadios, machos e fêmeas, os quais foram submetidos a colheita de sangue venoso por meio de punção da jugular externa. As amostras foram analisadas por meio de colorimetria manual. As análises foram feitas a cada 24 horas no intervalo de 14 dias. Durante este intervalo as amostras foram mantidas sob refrigeração. A variação da concentração de hemoglobina foi de 8,546 a 22,456 g/dL. Em comparação aos valores basais, não houve significativas alterações na concentração de hemoglobina durante o intervalo de tempo investigado, indicando que sua mensuração pode ser realizada em até 14 dias pós colheita.(AU)

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Hemoglobin is a protein present in erythrocytes, used in the transport of respiratory gases. This protein can be measured through laboratory tests, thus it is an important tool in health assessment of domestic animals. The hemoglobin test protocol by cyano-met hemoglobin method describes that its measurement can be made during a period of up to seven days, but the laboratorial routine shows a reduction in the values before this period. The objective of this study was to determine what the actual maximum period to measure hemoglobin levels in cattle is without changes. Nineteen healthy bovine animals were used, males and females, who underwent venous blood collection via jugular puncture. Samples were analyzed by manual colorimetric. Analyses were made every 24 hours during a fourteen days-interval. During this interval the samples were kept under refrigeration. Hemoglobin levels varied from 8.546 to 22.456 g/dL. Compared to baseline, there were no significant changes in hemoglobin concentration during the investigated period of time, indicating that its measurement can be performed after 14 days of storage.

A hemoglobina é uma proteína presente nos eritrócitos, que realiza o transporte dos gases respiratórios. Exames laboratoriais mensuram esta proteína, sendo uma importante variável para a avaliação de saúde em animais domésticos. O protocolo do teste da determinação da concentração de hemoglobina pelo método ciano-metemoglobina descreve que sua mensuração pode ser realizada em um prazo de até sete dias, porém a rotina laboratorial mostra redução nos valores antes do período proposto. O objetivo deste trabalho foi avaliar qual o real prazo máximo para mensurar as concentrações de hemoglobina em bovinos sem que haja alterações em seus valores. Foram utilizados 19 bovinos sadios, machos e fêmeas, os quais foram submetidos a colheita de sangue venoso por meio de punção da jugular externa. As amostras foram analisadas por meio de colorimetria manual. As análises foram feitas a cada 24 horas no intervalo de 14 dias. Durante este intervalo as amostras foram mantidas sob refrigeração. A variação da concentração de hemoglobina foi de 8,546 a 22,456 g/dL. Em comparação aos valores basais, não houve significativas alterações na concentração de hemoglobina durante o intervalo de tempo investigado, indicando que sua mensuração pode ser realizada em até 14 dias pós colheita.

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Background: The Flemish Red is one of the oldest breeds of French cattle and, in France, the crosses with other red breeds and replacing them with more productive breeds caused a severe reduction in the number of purebred Flemish Red animals. Due to this drastic decrease in the Flemish Red population, this breed is at risk of extinction. The objective of the present study is to describe follicular development and serum progesterone (P4) profiles in Flemish Red cows raised in Southern Brazil for the improvement of reproductive management of this breed, considering that these animals may exhibit traits which disappeared in European animals due to genetic dilution.Materials, Methods & Results: The estrus cycles of two groups of post-pubertal non-lactating Flemish Red (FR, n = 7) and Holstein (HOL, n = 7) cows were synchronized with a Prostaglandin F2α analog. From the day of estrus, the ovaries were evaluated by transrectal ultrasound every 24 h for 21 days or until the detection of the second ovulation. The diameters of the dominant follicle (DF) and subordinate follicles (SF) were recorded according to the day of the estrus cycles and analyzed based on the diameter of both DF and SF on the day of follicular emergence, the day and diameter of DF on the day of divergence, daily growth rates of DF and FS, and the maximum diameter of both the ovulatory follicle and the dominant non-ovulatory follicle (DNOF). Samples of blood were collected every five days for P4 measurement (RIA). During emergence of the first follicle wave, the DF measured 3.97 ± 0.19 mm for FR and 4.00 ± 0.35 mm for HOL, while the SF reached 3.40 ± 0.22 mm in FR and 3, 07 ± 0.26 mm in HOL. The daily follicle growth rates was 1.10 ± 0.04 mm for DF and 0.67 ± 0.06 mm for the SF in FR, and were 1.01 ± 0.05 mm for DF and 0.72 ± 0.09 mm for the SF in HOL.[...](AU)

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Background: The Flemish Red is one of the oldest breeds of French cattle and, in France, the crosses with other red breeds and replacing them with more productive breeds caused a severe reduction in the number of purebred Flemish Red animals. Due to this drastic decrease in the Flemish Red population, this breed is at risk of extinction. The objective of the present study is to describe follicular development and serum progesterone (P4) profiles in Flemish Red cows raised in Southern Brazil for the improvement of reproductive management of this breed, considering that these animals may exhibit traits which disappeared in European animals due to genetic dilution.Materials, Methods & Results: The estrus cycles of two groups of post-pubertal non-lactating Flemish Red (FR, n = 7) and Holstein (HOL, n = 7) cows were synchronized with a Prostaglandin F2α analog. From the day of estrus, the ovaries were evaluated by transrectal ultrasound every 24 h for 21 days or until the detection of the second ovulation. The diameters of the dominant follicle (DF) and subordinate follicles (SF) were recorded according to the day of the estrus cycles and analyzed based on the diameter of both DF and SF on the day of follicular emergence, the day and diameter of DF on the day of divergence, daily growth rates of DF and FS, and the maximum diameter of both the ovulatory follicle and the dominant non-ovulatory follicle (DNOF). Samples of blood were collected every five days for P4 measurement (RIA). During emergence of the first follicle wave, the DF measured 3.97 ± 0.19 mm for FR and 4.00 ± 0.35 mm for HOL, while the SF reached 3.40 ± 0.22 mm in FR and 3, 07 ± 0.26 mm in HOL. The daily follicle growth rates was 1.10 ± 0.04 mm for DF and 0.67 ± 0.06 mm for the SF in FR, and were 1.01 ± 0.05 mm for DF and 0.72 ± 0.09 mm for the SF in HOL.[...]

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