Effects of cell concentration during cryopreservation on the post-thaw quality of Santa ines ram sperm.

BACKGROUND: Non-surgical artificial insemination techniques for sheep may benefit from larger numbers of sperm in the insemination dose because the ewe cervix is convoluted and often cannot be traversed with an insemination gun resulting in deposition of the sperm at the os cervix. OBJECTIVE: To compare a range of sperm concentrations when cryopreserving semen from Santa Ines rams and determine the effects of this on post-thaw quality. MATERIALS AND METHODS: One ejaculate from each ram (n = 10) was diluted to four sperm concentrations to obtain the following groups: G-400, G-800, G-1200, and G-1600 x 106 sperm/mL. The semen samples were packaged in 0.25 mL straws, cooled to 5 degree C, cryopreserved in liquid nitrogen vapor, thawed in a water bath (40 degree C per 20 s), and were analyzed for computerized kinetics, capacitation and acrosome integrity, and plasma membrane integrity of sperm. RESULTS: The G-400 treatment resulted in samples with the highest linearity and progressive motion (P < 0.05) and had significantly greater plasma membrane integrity, and lower capacitation and acrosome reaction rates compared to G-1600 (P < 0.05). Overall, use of the G-400 treatment resulted in better kinetics, less plasma membrane damage and less early capacitation. However, despite reducing the ejaculate yield and increasing the costs of the semen freezing process, the G-800 and G-1200 treatments make a greater absolute number of sperm with good kinetics, plasma membrane integrity and capacitation status available. CONCLUSION: Ram sperm concentration impacts cryopreservation, and higher concentrations may be advantageous if a single artificial insemination protocol is desirable. doi.org/10.54680/fr22610110812.

Origin and population structure of Brazilian hair sheep breeds.

Brazilian hair sheep constitute a genetic diversity hotspot. These animals are found in the harsh environments of the Brazilian Northwest (semi-arid) region. Genotypes (50K SNP chip) from seven Brazilian sheep breeds (five hair and two coarse wool types) and 87 worldwide breeds were used to test for population structure, admixture and genetic diversity. Moreover, phylogenetic trees evaluating migration events between genetic groups were built. Brazilian Somali, a fat-tailed breed, had a close relationship with East African breeds and clustered distinctly from other Brazilian breeds. Brazilian Blackbelly and Barbados Blackbelly had a close relationship. The Morada Nova breed did not show close relationships with European or African breeds, revealing a single migration event from an Algerian hair breed. Brazilian Fat-tail and Morada Nova share a common ancestor, but the former showed introgressions from Brazilian Somali and Afrikaner breeds, explaining the fat-tail phenotype. The Santa Inês breed received a substantial contribution from Brazilian Bergamasca and showed an admixed origin with recent introgressions from other breeds, mainly from Suffolk. Furthermore, Brazilian Somali and Brazilian Fat-tail are the most endangered sheep genetic resources in Brazil and should be the focus for ex situ conservation programs. In conclusion, Brazilian hair sheep show an African origin and are characterized by diverse genetic composition, reinforcing the need for conservation of these genetic resources, and at the same time, this highly diverse group has variability that can be used in breeding programs.

Evaluation of morphology, morphometry and follicular dynamics in FecGE genotyped ewes.

This study aimed to evaluate the effect of FecGE mutation on the development of ovarian follicles. To this end, 42 Santa Inês ewes were genotyped for FecGE mutation and classified as wild-type (FecG+/+), heterozygous (FecG+/E) or mutant homozygous (FecGE/E). Ovarian fragments were processed, and the follicles were analyzed with regard to the morphology and morphometry using classical histology. For the evaluation of follicular dynamics, ewes underwent oestrous synchronization and were monitored throughout an interovulatory period. A higher (P < 0.05) percentage of morphologically normal follicles in the primordial stage was identified in FecGE/E (90.0%) and FecG+/E (88.1%) ewes than in the FecG+/+ (73.0%) ewes. There was also a significantly greater (P < 0.05) number of morphologically normal follicles in the FecGE/E (87.3%) and FecG+/E (83.3%) ewes than in FecG+/+ (76.8%) ewes in the transitional stage. A smaller (P < 0.05) diameter was observed in the secondary follicles in FecGE/E (93.8 µm) ewes than in FecG+/E (171.8 µm) ewes. Regarding follicular dynamics, FecGE/E ewes showed a greater (P < 0.05) number of ovulations (2.5 ±â€¯0.2) than FecG+/+ ewes (1.5 ±â€¯0.3) ewes. Ovulatory follicles were smaller (P < 0.05) in the FecGE/E (5.1 mm) and FecG+/E (5.2 mm) ewes than in FecG+/+ (5.8 mm) ewes. Santa Inês nulliparous ewes carrying the FecGE mutation showed a greater proportion of morphologically normal follicles in the primordial and transitional stages than those not carrying the mutation. FecGE/E ewes demonstrated a higher number of ovulated follicles and that FecGE/E and FecG+/E ewes presented ovulatory follicles with a smaller diameter.

Single nucleotide polymorphisms in the growth hormone and IGF type-1 (IGF1) genes associated with carcass traits in Santa Ines sheep.

Polymorphisms in the growth hormone (GH) and IGF type-1 (IGF1) genes have been associated with the economic traits in farm animals, including BW of some sheep breeds. However, it remains unknown if these polymorphisms also affect carcass traits in sheep. Thus, we aimed to identify polymorphisms in the GH and IGF1 genes in Santa Ines sheep in order to describe their allelic and genotypic frequencies as well as to test the hypotheses that they are associated with the carcass traits. Fragments of 4550 bp (IGF1) and 1194 bp (GH) were sequenced in up to 191 lambs. In all, 18 polymorphisms were identified in the IGF1 and 21 in the GH gene. The IGF1 polymorphisms rs430457475, rs412470350, rs409110739 and rs400113576 showed an additive effect on the internal carcass length (-0.9265±0.4223), rump girth (-2.9285±1.1473), rib yield (-1.0003±0.4588) and neck weight (-0.0567±0.0278), respectively. In addition, the polymorphisms rs58957314 in the GH affected the rib weight (-0.4380±0.1272) and rib yield (-2.2680±0.6970), loin weight (-0.1893±0.0516) and loin yield (-0.9423±0.3259), palette weight (-0.2265±0.0779) and palette yield (-0.9424±0.4184), leg weight (-0.3960±0.1375), neck weight (-0.0851±0.0394) and carcass finishing score (-0.1700±0.0839). These results allow us to conclude that there are polymorphisms in the IGF1 and GH genes associated with carcass traits in Santa Ines sheep, which can provide important information for marker-assisted selection.

Use of aqueous extract of noni in extender for sheep semen freezing / Utilização do extrato aquoso de noni em diluente para congelação de sêmen ovino

The study aimed to evaluate the action of aqueous extract of noni in an extender for sheep semen freezing. Treatments differed in inclusion of aqueous extract of noni in the extender: T1 ˗ no addition; T2 ˗ 24µg/mL; T3 ˗ 72µg/mL; and T4 ˗ 120µg/mL. Ejaculates were collected, diluted in the four treatments, and frozen. After thawing, the semen was subjected to a thermoresistance test and evaluated for subjective motility, vigor, membrane integrity assessment by hypo-osmotic swelling test, live-dead assay, computer-assisted sperm analysis and the status of sperm capacitation and acrosome reaction. Data were subjected to ANOVA, and then to Student Newman Keuls's test at 5% significance level. In the thermoresistance test after two hours of incubation, motility in T4 (120µg/mL) was lower than in the other treatments, with no differences in the HoS test in either diluted semen or in the semen evaluated immediately post-thawing, while for the other times, treatments showed similar responses. Regarding the motility parameters, a difference was observed for progressive motility, curvilinear velocity, average path velocity, and amplitude of lateral head displacement. As to the sperm capacitation status, a difference was observed between treatments for the sperm capacitated with intact acrosome.(AU)

Este estudo teve como objetivo avaliar a ação do extrato aquoso de noni em diluente para congelação de sêmen de carneiro. Os tratamentos diferiram quanto à inclusão de extrato aquoso de noni ao meio diluidor em: T1˗ sem adição de extrato; T2˗ 24µg/mL ; T3- 72µg/mL e 120µg/mL. Por meio de vagina artificial, 16 ejaculados foram coletados, diluídos entre os quatro tratamentos e congelados. Após o descongelamento, o sêmen foi submetido ao teste de termorresistência e avaliado quanto à motilidade subjetiva, ao vigor espermático, à integridade de membrana pelo teste hiposmótico, bem como ao teste supravital, à análise de sêmen assistida por computador (CASA) e ao status de capacitação espermática e de reação acrossomal. Os dados foram submetidos a uma análise de variância, seguida pelo teste de Student-Newman-Keuls com 5% de significância. No teste de termorresistência, após duas horas de incubação, a motilidade do T4 (120µg/mL) apresentou-se inferior aos demais tratamentos. Não houve diferença significativa no teste HOS tanto para o sêmen diluído quanto para o sêmen avaliado imediatamente pós-descongelação; para as demais horas, os tratamentos apresentaram comportamento semelhante. Para os parâmetros de cinética, foi observada diferença estatística para motilidade progressiva, velocidade curvilinear, velocidade do percurso médio e amplitude de deslocamento lateral da cabeça. Quanto ao estado de capacitação espermática, observou-se diferença entre os tratamentos para espermatozoide capacitado com acrossomo intacto.(AU)

Fish sperm subpopulations: Changes after cryopreservation process and relationship with fertilization success in tambaqui (Colossoma macropomum).

Fish tambaqui (Colossoma macropomum) is the native Brazilian fish with the highest agricultural production under intensive aquaculture in South America. However, the decrease in the genetic variability in fish farms has become necessary the improvement of cryopreservation process through new statistical studies of spermatozoa (like subpopulation studies). The evaluation of the kinetic data obtained with a computer-assisted sperm analysis system, applying a two-step cluster analysis, yielded in tambaqui three different subpopulations in fresh sperm: SP1, considered as a slow nonlinear subpopulation; SP2, considered as a fast nonlinear subpopulation, and finally; SP3, considered as a fast linear subpopulation. For cryopreserved sperm, the cluster analysis yielded only two sperm subpopulations: SP1', considered as a slow nonlinear subpopulation and SP2', which seemed to be an intermediate subpopulation (showing medium motility and velocity values) merged from SP2 and SP3 obtained from fresh sperm. Coefficients of correlation (r) and determination (r2) between the sperm subpopulations from fresh sperm and the fertilization rates were calculated, and SP2 and SP3 (the fast-spermatozoa subpopulations) showed a high-positive correlation with the fertilization rates (r = 0.93 and 0.79, respectively). In addition, the positive significant correlations found in curvilinear velocity (r = 0.78), straight line velocity (r = 0.57), and average velocity (r = 0.75) indicate that sperm kinetic features seem to be a key factor in the fertilization process in tambaqui, as occur in other fish species.

Cryopreservation of tambaqui (Colossoma macropomum) semen: extenders, cryoprotectants, dilution ratios and freezing methods.

The tambaqui is an Amazonian fish of great economic and environmental importance to Brazil and other South American countries. Several semen cryopreservation methodologies have been tested for different Brazilian fish species; however, there is little information on the use of this technique on tambaqui semen. The aim of the present study was to investigate the effect of osmolarity and activation solutions on sperm kinetics and, glucose solutions, cryoprotectants, dilution ratios, egg yolk and freezing methods on tambaqui semen freezing. The osmolarity of 230 mOsm was suitable for simultaneously yielding higher sperm motility (85%) and motility time (54 sec.) and osmolarities above 360 mOsm maintain immobile tambaqui sperm. The tambaqui semen can be successfully cryopreserved when diluted 1:9 in freezing medium composed of 5 percent glucose solution (290 mOsm) with 10 percent methylglycol and 5 percent egg yolk, and frozen directly in a dry shipper container.

Genetic diversity and assessment of 23 microsatellite markers for parentage testing of Santa Inês hair sheep in Brazil.

Santa Inês is the most common hair sheep breed in Brazil and probably has the highest genetic diversity among sheep breeds in this country. Successful breeding programs for Brazilian sheep breeds are not common for various reasons, including a lack of control of parentage in the flocks. We developed an allele frequency database for 23 STR loci for the Santa Inês breed based on 285 animals sampled from five populations distributed across the central-western and north-eastern regions of Brazil. The marker set included seven microsatellites used in the 2011 International Society for Animal Genetics sheep genotyping comparison tests and all eight microsatellites currently approved by the Brazilian Agricultural Ministry laboratory accreditation guidelines for sheep identification. The microsatellites had an average of 10 alleles and a mean expected heterozygosity of 0.745. Combined paternity exclusion probabilities when no parent or one parent was known were >99.99%. A small proportion (5.8%) of the existing genetic variation was found to be among the Santa Inês populations, possibly derived from genetic drift and selection. We found that the marker panel proposed by the Agricultural Ministry, although generally useful, should be enhanced by including more markers for improved exclusionary power in parentage testing. This database provides a useful tool for parentage testing of this major Brazilian breed, contributing to improved management and breeding of existing herds.

A new polymorphism in the Growth and Differentiation Factor 9 (GDF9) gene is associated with increased ovulation rate and prolificacy in homozygous sheep.

Brazilian Santa Inês (SI) sheep are very well-adapted to the tropical conditions of Brazil and are an important source of animal protein. A high rate of twin births was reported in some SI flocks. Growth and Differentiation Factor 9 (GDF9) and Bone Morphogenetic Protein 15 (BMP15) are the first two genes expressed by the oocyte to be associated with an increased ovulation rate in sheep. All GDF9 and BMP15 variants characterized, until now, present the same phenotype: the heterozygote ewes have an increased ovulation rate and the mutated homozygotes are sterile. In this study, we have found a new allele of GDF9, named FecG(E) (Embrapa), which leads to a substitution of a phenylalanine with a cysteine in a conservative position of the mature peptide. Homozygote ewes presenting the FecG(E) allele have shown an increase in their ovulation rate (82%) and prolificacy (58%). This new phenotype can be very useful in better understanding the genetic control of follicular development; the mechanisms involved in the control of ovulation rate in mammals; and for the improvement of sheep production.

Avaliação de uma metodologia profilática contra a mastite clínica em ovelhas da raça Santa Inês / Evaluation of a prophylactic method against clinical mastitis in Santa Inês ewes

Mastitis is an important problem for the Santa Inês ewes and is a disease strongly related to the management carried out in the herd. A comparative study was carried out between the proposed method based on drying (treatment) and the traditional method (control). Sixty-two Santa Inês ewes raised in a semi-extensive system were divided in two groups of 31 ewes each: treatment group (G1) and control group (G2). The animals were monitored each week searching for clinical sings of mastitis. In 8.4 percent of G1 and 41.3 percent of G2 animals, the udder was warm and hyperemic. The presence of residual milk was lower in G1 (24.3 percent) compared to those of G2 (74.2 percent). The proposed treatment for the control of ovine mastitis significantly reduced the frequency of the clinical signs of the disease.

Induçäo do parto em cabras de raças leiteiras mediante aplicaçäo de cloprostenol / Induction of parturition in dairy goat by administration of cloprostenol

Conduziram-se dois experimentos com o objetivo de avaliar a eficiência do cloprostenol em diferentes doses e vias de aplicaçäo na induçäo do parto em cabras. No primeiro, 14 cabras das raças leiteiras Anglo-Nubiana, Saanen e Pardo-Alpina foram submetidas, no 145º dia de prenhez, a três tratamentos: T1.1 = 25µg (n=04), T1.2 = 50µg (n=05) e T1.3 - 100µg (n=05) de cloprostenol, administrados por via intramuscular na vulva (IMV). Três cabras prenhes das mesmas raças foram utilizadas como controle. Os partos ocorreram com: T1.1 = 30,13ñ4,77h, T1.2 - 32,43ñ4,82h e T1.3 = 34,29ñ3,96h, e a amplitude do tempo de ocorrência dos partos foram de: 11,58h, 12,25h e 7,40h, respectivamente. Näo foram observadas diferenças estatísticas entre os tratamentos, embora T1.3 tenha apresentado uma resposta mais consistente (CV=11,55 por cento) que T1.1 (CV=15,83 por cento) e T1.2 (CV=14,86 por cento). No segundo experimento, 51 fêmeas das mesmas raças foram distribuídas, no 145º dia de prenhez, em quatro tratamentos: T2.1 = 25µg (n=14), T2.2= 50µg (n=12), T2.3 = 75µg (n=12) de cloprostenol administradas IMV e T2.4 - 75µg (n=13) de cloprostenol na musculatura da coxa (IM). Os intervalos entre a induçäo e o parto foram de: T2.1 = 34,89ñ13,57h, T2.2 = 30,06ñ3,82h, T2.3 = 29,99ñ1,81h e T2.4 = 37,44ñ22,15h. Näo houve diferença estatística significativa entre os tratamentos. Entretanto, T2.3 mostrou-se mais consistente ao apresentar coeficiente da variaçäo menor (CV=6,04 por cento) que T2.1 (CV=38,88 por cento), T2.2 (CV=12,70 por cento) e T2.4 (CV=59,17 por cento). Constatou-se, também, que os tratamentos permitiram a concentraçäo dos partos no 146º dia de prenhez, com amplitude de ocorrência de 0,62 dia e 3,07 dias no primeiro e segundo experimento, respectivamente, contra 11 dias para o grupo controle no primeiro experimento. Conclui-se que as doses de 25, 50 e 75µg de cloprostenol, aplicadas por via IMV, foram efetivas em induzir o parto em cabras, sendo a dose de 75µg aparentemente a mais eficaz

Efeito da insulaçäo escrotal sobre os constituintes do plasma seminal de bodes / Effect of scrotal insulation on seminal plasma of bucks

No plasma seminal de bodes da raça Moxotó e ½ sangue Moxotó-Pardo Alpina foram quantificadas a concentraçäo de frutose, ácido cítrico e proteína total, além das enzimas aspartato aminotransferase (AST) e alanina aminotransferase (ALT), em espectrofotometria, com comprimento de onda de 505 mm, antes e após aquecimento do saco escrotal. A insulaçäo do saco escrotal com bolsa plástica, de parede dupla, durante, 6,5 dias, provocou aumento expressivo na concentraçäo da frutose seminal, inversamente proporcional à concentraçäo espermática, só retornando à normalidade com a regeneraçäo do quadro espermático. A concentraçäo do ácido cítrico e da proteína total näo apresentou diferenças estatísticas significativas entre grupos genéticos, diante do desafio calórico. As concentraçöes da AST e ALT reduziram-se pela metade após o desafio térmico até o nono período (semana) e só retornaram à normalidade a partir do 10º período experimental. Dez semanas após o início do desafio escroto-testicular todos os animais mostraram regeneraçäo dos níveis de frutose, ácido cítrico, proteína total e das enzimas AST e ALT no plasma seminal

Eficiência do flunixin meglumine no controle da regressäo lútea prematura em cabras superovuladas / Efficiency of flunixin meglumine in the control of premature luteal regression in supervovulated goats

Testou-se a eficácia de três doses de flunixin meglumine, um anti-prostaglandínico que impede a regressäo lútea precoce, em 32 cabras, sob estro sincronizado com esponjas vaginais, por 11 dias, impregnadas com 60mg de acetato de medroxiprogesterona, e aplicaçäo intramuscular de 50µg de cloprostenol no nono dia da sincronizaçäo. Nesse dia, também, teve início a superovulaçäo das doadoras com 9mg de FSH, fracionadas em seis aplicaçöes, em doses decrescentes, com intervalo de 12 horas. Três dias após a retirada das esponjas, iniciaram-se os tratamentos (T) com flunixin meglumine, nas seguintes doses por aplicaçäo: T1=0,00; T2=1,10; T3=1,65 e T4=2,20mg/kg de peso vivo, em um esquema de oito aplicaçöes, intervaladas de 12 horas. As fêmeas foram cobertas por reprodutores de fertilidade comprovada e as colheitas dos embriöes foram realizadas por laparotomia, entre o sexto e oitavo dia após a última cobertura. Os sintomas de estro foram observados em 84,34 por cento das fêmeas após os tratamentos de sincronizaçäo e superovulaçäo, principalmente entre 12 e 36 horas da retirada das esponjas. A ovulaçäo ocorreu em 87,10 por cento dos animais. As porcentagens de corpos lúteos funcionais em T1, T2, T3 e T4 foram, respectivamente, 54,95 e 45,04; 98,06 e 1,94; 67,16 e 32,83; 80,41 e 19,58. As maiores porcentagens de recuperaçäo embrionária ocorreram em T2; 51,45 e T4; 49,48 e as menores em T3 e T1, 32,83 e 28,83, respectivamente, mostrando relaçäo positiva com os porcentuais de corpos lúteos funcionais. T2 e T1 apresentaram os maiores valores de embriöes viáveis, 64,15 por cento e 56,25 por cento, respectivamente, seguidos por T4, 45,83 por cento e T3, 36,36 por cento. A dose de 1,10mg/kg de flunixin meglumine apresentou melhores resultados

Creutzfeldt-Jakob disease. A survey of 14 patients.

Creutzfeldt-Jakob disease (CJD) is a transmissible disease of the nervous system causatively related to the presence of an abnormal prion protein, with dementia, myoclonic jerks, and periodic EEG activity. Fourteen patients (7 females and 7 males) ranging from 26 to 76 years of age (median 59 years) were evaluated between 1974 and 1995 at the Neurologic Clinic of São Paulo University School of Medicine. The average duration of the disease was 12 months (3.5-34 months). Early clinical findings were: behaviour changes in 7 patients, dementia in 4, visual disturbances in 4, vertigo in 2, tremor in 9, and dystonia in one. Advanced symptoms were dementia and myoclonus in all patients. Pyramidal tract dysfunction was found in 6, cerebellar ataxia in 2, seizures in 3, nystagmus and vertigo in 4, and peripheral nervous system involvement in 2. Atypical clinical forms were found in 5 patients. Periodic EEG activity was found in 10 patients. Cerebrospinal fluid evaluation showed pleocytosis in 1 patient, higher protein content in 2, and higher gamma globulin level in 2. In 10 patients anatomopathological evidence in the central nervous system confirmed the clinical diagnosis by presenting with status spongiosus. All except one patient presented with the sporadic form of the disease.

[Congenital myotonia. Report of 7 patients]. / Miotonia congênita. Relato de sete pacientes.

Myotonia is the phenomenon of decrease of muscular relaxation rate, after either a contraction or a mechanical or electrical stimulus. Congenital myotonias are hereditary affections and do not present muscular dystrophy. The current trend is to group them as ionic channels diseases, together with the periodic paralysis. The authors accompanied the cases of seven patients, six males and one female, with ages ranging from 16 to 48 years (average 27 years) and onset of symptoms between 1 and 10 years (average 5 years). These patients presented a myotonic phenomenon unleashed by intensive contraction and global muscular hypertrophy. Three patients were diagnosed as cases of Becker type generalized myotonia because they presented a recessive autosomic heredity and/or transient episodes of muscular weakness. Two patients fitted the description of Thomsen congenital myotonia, with a pattern of dominating autosomic heredity and/or absence of weakness episodes or worsening factors for their condition. Two patients presented fluctuating myotonia, which because worse in cold weather or at potassium intake. The clinical diagnosis was confirmed through complementary tests (electroneuromyography, muscle biopsy and DNA study). Each of the patients made use of different drugs, in the search of optimal lessening of their myotonia. There were five reports of amelioration with the use of diphenilhydantoine; one report with the use of carbamazepine; three reports with the use of acetazolamide; one report with the use of a calcium channel blocker; one report with the use of a beta-adrenergic; one report with the use of thiazide; and none with the use of quinidine/procainamide.

[Nemalinic myopathy with intracytoplasmic spheroid bodies: report of a case]. / Miopatia nemalínica com corpos intracitoplasmáticos esferóides: relato de caso.

The authors report the case of a female patient, 18 years of age, with slowly progressing weakness in upper and lower limbs since childhood. There were no significant antecedents. The neurologic examination showed mild proximal and distal motor deficit with a slight muscular retraction at the level of shoulders, elbows, coxofemural joints, knees and ankles; muscular hypotrophy in the legs and feet; reflexes were present and sensitivity was normal. Creatinephosphokinase showed an increase of one and a half times the normal value. Electroneuromyography: decrease in the amplitude and duration of action potentials and excessive recruitment of motor units, compatible with a primary muscular disease. A muscle biopsy with frozen sections (HE, Gomori, PAS, ATPases, NADH, SDH, acid and alcaline phosphatases, cytochrome oxidase and Oil-red-o) revealed a primary muscular disease characterized by the presence of nemalinic and intracytoplasmic spheroid bodies. Nemalinic bodies have been described with different structural abnormalities of muscle fibers; however, such association is rare. This is the second case report of concomitant occurrence of nemalinic and spheroid bodies.