Canine pulmonary clearance during feto-neonatal transition according to the type of delivery.

The success of immediate adaptation to extrauterine life depends on appropriate lung function, however, elective cesarean section can increase the risk of respiratory distress as a result of reduced pulmonary fluid absorption. This study aimed to evaluate the influence of birth mode on pulmonary clearance and respiratory performance of canine neonates in the transition period. For this purpose, 37 neonates were selected according to the obstetric condition: Vaginal Eutocia (n = 17) and Elective C-section (n = 20). Neonates were evaluated for neonatal vitality score, as well as evaluation of heart and respiratory rates, body temperature and body weight, venous hemogasometric evaluation, blood lactate and glucose, pulse oximetry and radiographic evaluation during the first 24 h of life. Additionally, amniotic fluid electrolyte composition of each puppy was evaluated. There was no influence of the type of delivery on electrolyte composition of canine amniotic fluid and neonatal pulmonary liquid content, analyzed by thoracic X-Rays. On the other hand, elective cesarean section delayed pulmonary adaptation, resulting in hypoxemia and less efficient compensatory response to acid-base imbalance and thermoregulation. In conclusion, elective c-section does not delay pulmonary clearance, whilst alters pulmonary adaptation by less efficient gas exchange and lower oxygenation, hindering the compensatory response to acid-base imbalance during the fetal-neonatal transition in dogs.

Frozen-Thawed Sperm Analysis of Benign Prostatic Hyperplasia Dogs Treated With Finasteride.

Benign Prostatic Hyperplasia (BPH) is a pathological condition that directly interferes with the reproductive potential of senile dogs, by leading to prostate enlargement and sperm injury, which in turn may compromise sperm freezeability. Moreover, albeit finasteride treatment reduces prostatic volume and blood supply and maintains seminal quality and testicular integrity, the effects of sperm samples submitted to cryopreservation after the finasteride treatment are still unknown. Thus, the aim of this study was to evaluate frozen-thawed semen of BPH dogs, as well as dogs subjected to BPH pharmacological treatment with finasteride. For such purpose, 20 dogs were previously selected and assigned to three experimental groups, according to BPH diagnosis and treatment with finasteride: Control (n = 9), BPH Group (n = 5) and BPH-Finasteride Group (n = 6). Semen was subjected to one-step cryopreservation protocol with tris-fructose-citric acid extender with 5% glycerol and thawed at 37°C for 30 sec. Fresh and post-thaw sperm samples were evaluated for macroscopic parameters, sperm concentration, sperm motility kinetics, sperm mitochondrial activity and potential, oxidative stress, plasmatic and acrosome membrane integrity, sperm DNA fragmentation and sperm binding test on perivitelic membrane of chicken egg yolk. Regarding fresh semen, BPH-Finasteride group had the lowest ejaculate visual aspect (opacity), higher frequency of sperm flagellar beating (BCF) and percentage of sperm with medium velocity. Control group had the highest percentage of sperm DNA integrity compared to BPH group. For the frozen-thawed semen, Control group presented the highest percentage of spermatozoa with high mitochondrial activity. However, the BPH-Finasteride group showed higher number of sperm bound to the perivitelline membrane of chicken egg yolk compared to the BPH Group. Conversely, BPH group had higher percentage of DNA damage. In conclusion, the ejaculate of BPH dogs has higher susceptibility to cryoinjury, whereas finasteride-treated dogs have increased spermatozoa functional performance, suggesting a promising use of BPH dogs as semen donors in sperm cryopreservation programs.

A triple stain method in conjunction with an in-depth screening of cryopreservation effects on post-thaw sperm in dogs.

In order to accurately analyze the possible side effects of sperm cryopreservation, an in-depth screening of post-thaw sperm status is necessary. Thus, this study aimed to identify thorough effects of sperm cryopreservation, by evaluating the integrity of all specific structures of the canine spermatozoa. Thirteen (n = 13) mature dogs of different breeds were selected. Six dogs (n = 6) were subjected to sperm cryopreservation, whereas seven dogs (n = 7) were used as semen donors to validate a simultaneous assessment of sperm plasmatic, acrosomal, and mitochondrial membranes (triple stain) by fluorescent probes. Fresh and post-thaw semen samples were evaluated through a computer-assisted analysis of sperm motility, sperm morpho-functional evaluation, triple stain and sperm DNA integrity. Post-thaw semen samples had lower total and progressive motility, as well as higher percentage of minor and major defects. Moreover, post-thaw samples had higher percentage of sperm with plasma membrane and mitochondrial damage but intact acrosome, and also sperm with simultaneous damaged plasma, acrosomal and mitochondrial membranes. Furthermore, post-thaw sperm had higher protamination deficiency and DNA fragmentation. In conclusion, cryopreservation has a broad impact in sperm morphology and function, altering motility patterns, plasma, acrosome and mitochondrial membranes integrity, as well as sperm DNA.

Short-term cold storage of seminal samples of the golden lancehead pitviper (Bothrops insularis).

Assisted reproductive technologies such as cooling, freezing, and artificial insemination are essential in conserving genetic diversity of critically endangered reptiles like the golden lancehead pitviper (Bothrops insularis). Therefore, we examined viability of semen samples from captive golden lanceheads diluted in coconut-water based extender over a 48-h period of cooling (5°C). Semen evaluations were performed immediately after the dilution and at 6, 24, and 48 h, using computer-assisted sperm analysis and stains to assess plasma membrane and acrosomal status. Our findings showed that the extender and protocol employed here were effective in preserving golden lancehead pitviper spermatozoa for a short periods of time (48 h), allowing semen samples to be transported to distant locations for immediate use without the setbacks of cryopreservation.

Surgical correction of bilateral ectopic ureter using the neoureterocystostomy technique / Correção cirúrgica de ureter ectópico bilateral utilizando a técnica de neoureterocistostomia

Ureteral ectopy is a rare disorder in the small animals' clinic. It is characterized as a congenital anomaly, resulting from the ducts differentiation failure during embryogenesis. In this scenario, the ureters present themselves outside the anatomical site, being inserted into the uterus, urethra, urinary vesicle neck, or vagina. The clinical signs are urinary incontinence and perivulvar dermatitis. Surgery is the accepted treatment to correct the anomaly. The surgical procedure is based on relocating the ectopic ureter and treating associated modifications. This report describes a case of intramural bilateral ureteral ectopy, corrected surgically through the neoureterocystostomy technique, making it possible to control the animal's urinary incontinence.(AU)

A ectopia ureteral é uma afecção de incidência rara na clínica de pequenos animais, sendo caracterizada como anomalia congênita resultante de falha na diferenciação dos ductos durante a embriogênese. Neste cenário, os ureteres se apresentam fora do seu local anatômico, sendo inseridos no útero, no colo da vesícula urinária, na uretra ou na vagina. Os sinais clínicos comumente apresentados são a incontinência urinária bem como a dermatite perivulvar. O tratamento de eleição para correção da anomalia é o procedimento cirúrgico, no qual a técnica de escolha é baseada na relocação do ureter ectópico e tratamento das alterações associadas. Neste contexto, o presente relato descreve um caso de ectopia ureteral bilateral intramural, corrigido cirurgicamente por meio da técnica neoureterocistostomia, o que possibilitou controle da incontinência urinária do paciente.(AU)

Short-term cold storage of seminal samples of the golden lancehead pitviper (Bothrops insularis)

Assisted reproductive technologies such as cooling, freezing, and artificial insemination are essential in conserving genetic diversity of critically endangered reptiles like the golden lancehead pitviper (Bothrops insularis). Therefore, we examined viability of semen samples from captive golden lanceheads diluted in coconut‐water based extender over a 48‐h period of cooling (5°C). Semen evaluations were performed immediately after the dilution and at 6, 24, and 48 h, using computer‐assisted sperm analysis and stains to assess plasma membrane and acrosomal status. Our findings showed that the extender and protocol employed here were effective in preserving golden lancehead pitviper spermatozoa for a short periods of time (48 h), allowing semen samples to be transported to distant locations for immediate use without the setbacks of cryopreservation.

Effects of clamping umbilical cord on the neonatal viability of puppies delivered by cesarean section.

In human neonates, when the umbilical cord is kept intact postpartum, blood continues to flow to the neonate, but this procedure might be difficult in dogs owing to a shorter umbilical cord and several neonates in a litter. However, it might be possible to detach the placenta and keep the umbilical cord intact, allowing residual blood to flow to the puppies. This study compared the effects of clamping versus no clamping of the umbilical cord in dogs born by cesarean section on neonatal vitality. The puppies were assessed by Apgar and reflex scores. Fifty puppies delivered from 16 bitches were randomly allocated to receive immediate umbilical cord clamping (n=25) or no clamping for at least 3 min after the onset of breathing (n=25). The puppies were assessed during the first 5 min of life and 10 min after the first assessment. The no clamping group showed significantly higher Apgar scores (second assessment, P<0.01) and reflex scores (first and second assessments, P<0.05) than the clamping group, implying higher vitality in the no clamping group. The differences observed between the moments (first versus second assessment) of each group was significant (P<0.01), revealing higher vitality in the second assessment. The results suggest that keeping the umbilical cord intact for at least 3 min after the onset of breathing may contribute to increased vitality in puppies delivered by cesarean section without any negative consequences.

Effect of senescence on morphological, functional and oxidative features of fresh and cryopreserved canine sperm.

The present research aimed to compare the hormonal profile, sperm quality and freezability of young and senile dogs. Dogs were assigned into Young Group (n = 11) and Senile Group (n = 11), additionally divided into Fresh Semen Group and Cryopreserved Semen Group. Males were evaluated for libido score and blood estrogen and testosterone assay. Sperm morphofunctional evaluations were performed based on Computer Assisted Sperm Analysis, morphology, mitochondrial activity, mitochondrial membrane potential, plasma and acrosomal membrane integrity, and DNA fragmentation. Sperm oxidative features were: protein oxidation, lipid peroxidation and production of advanced glycation end-products. Young dogs had higher libido score, sperm velocity average pathway, linearity of motility and mitochondrial activity index and lower percentage of major defects, total defects and proximal cytoplasmic droplet, despite the lack of difference between hormone profile of aged dogs. Fresh semen of senile dogs had increased percentage of spermatozoa with high mitochondrial membrane potential compared to young dogs and to cryopreserved sperm. Cryopreserved semen of young dogs had higher acrosomal membrane integrity compared to the Senile Group. In conclusion, sperm of aged dogs have reduced quality, signaled by higher morphological defects, ultimately altering sperm mitochondrial function and sperm kinetics. Furthermore, spermatozoa from senile dogs are more sensible to cryoinjury.

Uterine intussusception in immediate postpartum in bitches: case report / Intussuscepção uterina no puerpério imediato em cadelas: relato de caso

Uterine intussusception is a rare condition in bitches. This study aimed to report an unusual case of prolapsed uterine intussusception in a female dog. A 2-year-old American Bully bitch was received with labor history of large fetus requiring manual traction. The animal presented mucosal prolapse through the vulva, unproductive contractions and abdominal discomfort. After partial correction of mucosal prolapse, the dog was referred to ovary-hysterectomy surgery to correct prolapsed mucosa. During the surgical procedure, surgeons observed that the uterine horns were invaginated into the uterine body, rotated, and the tissue was slightly devitalized and congested. As previously described, this condition occurred during the immediate postpartum period. Thus, we suggest that this period can be a facilitator for the establishment of uterine intussusception.(AU)

A intussuscepção uterina é uma condição rara em cadelas. Portanto, este estudo tem por objetivo relatar um caso incomum de intussuscepção uterina prolapsada em fêmea canina. Uma cadela American Bully de dois anos de idade com histórico de parto de feto absoluto grande no qual necessitou de auxílio por tração manual foi apresentada. O animal exibiu um prolapso de mucosa através da vulva, contrações improdutivas e dores abdominais. Após a retração parcial do prolapso de mucosa, a cadela foi encaminhada para a ovário-histerectomia que teve por objetivo, também, corrigir a mucosa prolapsada. Durante o procedimento cirúrgico, foram verificados os cornos uterinos invaginados no corpo uterino, rotacionados, com o tecido levemente desvitalizado e congesto. Assim, como poucos casos encontrados na literatura, esta condição ocorreu durante o puerpério imediato. Dessa forma, sugere-se que este período, embora não seja um fator determinante para o estabelecimento da intussuscepção uterina, pode ser um facilitador no estabelecimento dessa condição.(AU)

Spermatic mitochondria: role in oxidative homeostasis, sperm function and possible tools for their assessment.

SummaryDespite sperm mitochondrial relevance to the fertilization capacity, the processes involved in the production of ATP and functional dynamics of sperm mitochondria are not fully understood. One of these processes is the paradox involved between function and formation of reactive oxygen species performed by the organelle. Therefore, this review aimed to provide data on the role of sperm mitochondria in oxidative homeostasis and functionality as well the tools to assess sperm mitochondrial function.

Carnosine as malondialdehyde scavenger in stallion seminal plasma and its role in sperm function and oxidative status.

Semen biotechniques may impair sperm quality due to excessive production of reactive oxygen species (ROS). Additionally, products of the oxidative reaction, especially involving lipids (e.g., malondialdehyde - MDA), may be even more harmful to sperm. Carnosine, previously reported to be present in seminal plasma of several species, may be a key factor on sperm tolerance to biotechniques by counterattacking the deleterious influence of MDA. Therefore, the aim of this study was to measure the levels of carnosine present in equine seminal plasma and relate these findings with sperm function and oxidative status during cooling and cryopreservation. Thus, semen samples were collected from 40 stallions in duplicate (N = 80) and then submitted to cooling and cryopreservation. Samples were then allocated into groups of high and low tolerance to refrigeration and cryopreservation (bad cooler and good cooler/bad freezer and good freezer, respectively), and in groups of different concentrations of carnosine (High, Medium-high, Medium-low and Low carnosine). Samples were evaluated for sperm kinetics patterns, function of sperm structures and oxidative status. In good cooler samples, it was observed higher concentrations of carnosine (Good cooler: 224.98 ±â€¯19.16 ng/mL; Bad cooler: 159.72 ±â€¯15.99 ng/mL; p = 0.0056), ROS production (Good cooler: 26.40 ±â€¯18.33%; Bad cooler: 18.33 ±â€¯1.84%; p = 0.001) and lipid peroxidation rates (Good cooler: 193.23 ±â€¯18.22 ng/mL; Bad cooler: 131.92 ±â€¯12.25; p = 0.0064). Groups of samples with higher carnosine concentrations had lower levels of malondialdehyde (High: 79.33 ±â€¯6.72 ng/mL; Medium-high: 140.45 ±â€¯11.70 ng/mL; Medium-low: 202.57 ±â€¯16.30 ng/mL and Low: 231.02 ±â€¯32.35 ng/mL; p < 0.05), demonstrating that carnosine was effective in removing lipid peroxidation products. Due to the removal of seminal plasma during the cryopreservation process, no differences occurred in carnosine levels between bad and good freezer groups. In this context, this study provides relevant data for future therapies using carnosine during cryopreservation, aiming to replace the levels lost due to the necessary removal of seminal plasma.

Induced sperm oxidative stress in dogs: Susceptibility against different reactive oxygen species and protective role of seminal plasma.

Oxidative stress (OS) is characterized by an unbalance between increased levels of reactive oxygen species (ROS) and/or impaired antioxidant protection. In this context, the composition of seminal plasma (SP) plays a key role in protecting sperm against OS. However, reproductive biotechnologies applied to dogs recommend the removal of SP. Thus, antioxidant therapy may be an important alternative when applying biotechniques such as semen cryopreservation in this specie. However, in order to be efficient, the choice of the ideal antioxidant in each condition is essential since each ROS is preferably neutralized by different antioxidant systems. Therefore, this study aims to evaluate the susceptibility of canine spermatozoa to different oxidative challenges (superoxide anion [O2-], hydrogen peroxide [H2O2], hydroxyl radical [OH-] and malondialdehyde [MDA]) in the present or absence of SP. We used ejaculates of eight dogs and submitted to induce oxidative challenges (with or without SP). After incubations, samples were evaluated for the susceptibility to lipid peroxidation, motility, mitochondrial activity and function, DNA integrity, plasma membrane and acrosome integrity. Sperm with SP had mitochondrial function preserved against ROS. However, in the absence of SP, H2O2 reduced mitochondrial membrane potential. In addition, regardless on SP, H2O2 was deleterious to sperm kinetics and plasma/acrosomal membranes. Incubation with OH- reduced mitochondrial activity and increased DNA fragmentation also independent on the absence of presence of SP. Furthermore, samples with SP were more resistant to lipid peroxidation (i.e., decreased concentration of TBARS). In conclusion, H2O2 and OH- appears to be the most deleterious ROS to dog sperm and SP protects the spermatozoa against mitochondrial injuries and lipid peroxidation.

Effects of Soy Lecithin Extender on Dog Sperm Cryopreservation.

Semen cryopreservation is an essential biotechnology in canine reproduction and during the cryopreservation process commonly egg yolk are used. The discrepancy in the egg yolk composition and the potential risk of disease dissemination are obstacles for semen exportation and use. Therefore, studies aiming to substitute egg yolk are extremely important. In this context, soy lecithin contains a low-density lipoprotein fraction, is an interesting alternative. Thus, the objective of this study was to compare extenders based on soy lecithin (several concentrations and forms) with egg yolk during the cryopreservation process of dog sperm. For this purpose, we used twelve dogs. Semen was evaluated at different time points (after refrigeration, glycerolization, and thawing), by motility analysis (CASA) and functional tests (e.g., membrane integrity-eosin/nigrosin, acrosome integrity-fast green/Bengal rose, mitochondrial activity-3'3 diaminobenzidine, Chromatin susceptibility to acid-induced denaturation-SCSA, and susceptibility to oxidative stress-thiobarbituric acid reactive substances). The results indicated that egg yolk and lower concentrations of lecithin had similar effects on mitochondrial activity and motility. Thus, soy lecithin is a potentially viable alternative to egg yolk for the cryopreservation of dog semen.

Sperm-binding to the perivitelline membrane of chicken egg yolk as a functional test for sperm evaluation in dogs / Teste de ligação espermática à membrana perivitelínica da gema de ovo de galinha como avaliação funcional do sêmen de cães

During fertilization, spermatozoa interact with the zona pellucida (ZP) through the binding between the acrosome and proteins 2 and 3 (ZP2 and ZP3). The perivitelline membrane of chicken egg yolk is homologous to the mammalian ZP3, which allows the binding of sperm of several species. The aim of this study was to standardize and evaluate the efficiency of sperm-binding to the perivitelline membrane of chicken eggs as a functional method for canine semen evaluation. For this purpose, nine post-thaw sperm samples were used, which were divided into two aliquots: the first was kept in water bath at 37ºC (live sample) and the second was submitted to cold shock to induce cellular damage (dead sample). The two aliquots were mixed on five proportions, corresponding to 0%, 25%, 50%, 75%, and 100% of viable cells, and the binding test was performed by analyzing the number of spermatozoa bonded to the perivitelline membrane by means of computerized assessment of sperm motility (CASA) or conventional microscopy. Additionally, samples were submitted to sperm motility analysis, evaluation of plasmatic and acrosomal membrane integrity, and sperm mitochondrial activity. The sperm-binding test to the perivitelline membrane of chicken egg yolk was considered a feasible sperm analysis test for both fertilizing capacity and overall sperm attributes evaluation, mainly when the analysis is performed by a conventional microscope, which expands its practicality to the majority of canine reproduction laboratories.(AU)

Durante a fecundação, os espermatozoides interagem com a zona pelúcida (ZP) por meio da ligação entre o acrossomo e as proteínas 2 e 3 (ZP2 e ZP3). A membrana perivitelínica da gema de ovo de galinhas é homóloga à ZP3 de mamíferos, possibilitando a ligação espermática de diversas espécies. Este trabalho padronizou e avaliou a eficiência do teste de ligação espermática à membrana perivitelínica da gema de ovo de galinhas como avaliação funcional do sêmen de cães. Para tal, foram utilizadas nove amostras seminais previamente criopreservadas. Cada amostra foi dividida em duas alíquotas: a primeira foi mantida em banho-maria à 37ºC (vivos) e a segunda submetida a choque térmico com o intuito de induzir dano celular (mortos). As duas alíquotas foram misturadas, correspondendo a 0, 25, 50, 75 e 100% de células viáveis. As amostras foram avaliadas quanto ao número de espermatozoides ligados à membrana perivitelínica por meio da análise computadorizada da motilidade (CASA) ou microscopia convencional. Ademais, as amostras foram avaliadas quanto à motilidade espermática, integridade das membranas acrossomal e plasmática e atividade mitocondrial espermática. O teste de ligação espermática à membrana perivitelínica de ovos de galinha foi considerado um teste de análise seminal exequível tanto para avaliar a capacidade fecundante dos espermatozoides como atributos seminais gerais, especialmente quando realizado em microscopia convencional, expandindo sua praticidade para a maioria dos laboratórios de análise de sêmen canino.(AU)

Infertility diagnosis in jaguar (Panthera onca): case report / Diagnóstico de infertilidade em onça pintada (Panthera onca): relato de caso

This work reports one case of infertility in a male jaguar (Panthera onca) aged 21 years and weighing 125 kg. Changes in sperm due to chronic stress, inadequate food handling and reproductive senescence are emphasized.(AU)

O presente trabalho relata um caso de infertilidade em uma onça-pintada (Panthera onca), macho, de 21 anos e 125 kg. São ressaltadas as alterações ocorridas nos espermatozoides, devido a estresse crônico, manejo alimentar inadequado e senescência reprodutiva.(AU)

Pontos críticos no manejo pré-abate de frango de corte: jejum, captura, carregamento, transporte e tempo de espera no abatedouro / Critical points in the pre-slaughter management of broiler: feed withdrawal, capture and catching, transport and lairage time in the plant

Atualmente, a produção avícola brasileira configura o Brasil como o terceiro maior produtor mundial de carne de frango. A cadeia avícola é um setor extremamente organizado em todos seus segmentos, do nascimento ao abate das aves. Um dos procedimentos mais laboriosos é o período pré-abate, que compreende desde o jejum até o tempo de espera no abatedouro. Devido a esse fato, este trabalho salienta os principais pontos críticos existentes no período pré-abate: o jejum, a captura, o carregamento, o transporte e tempo de espera no abatedouro.

Today the brazilian poultry industry sets Brazil as the third largest producer of chicken meat. The poultry production sector is extremely organized in all its segments, from birth to slaughter poultry. One of the most laborious is the pre-slaughter, which ranges from the feed withdrawal to the lairage time at the plant. Due to this fact this research highlights the most critical points in the pre-slaughter period, as: feed withdrawal, capture, catching, transport and lairage time in the plant.