Abstract This study aimed to analyze the characteristics of the HSP70 gene and protein in spermatozoa of Bali bulls of different age groups and to examine its potential as a biomarker determining bull fertility. This study used frozen semen produced from six Bali bulls divided into two groups based on age (≤ 9 years and ≥ 12 years). Parameters of frozen semen quality analyzed included sperm motility and kinetics using computer-assisted semen analysis, sperm morphological defects using Diff-Quick staining, acrosome integrity using FITC-PNA staining, and DNA fragmentation using acridine orange staining. HSP70 gene expression characterization was analyzed using qRT-PCR, and HSP70 protein abundance was analyzed using enzyme immunoassays. Fertility field data were obtained by analyzing the percentage conception rate for each bull based on the artificial insemination service data contained in the Indonesian-integrated system of the National Animal Health Information System (iSIKHNAS). The results showed significant differences (P<0.05) in total and progressive motility, morphological defects of the neck and midpiece, and tail of sperm, and acrosome integrity between the age groups of Bali bulls. HSP70 gene expression and protein abundance showed no significant differences (P>0.05) in different age groups. HSP70 gene expression correlated with fertility rate (P<0.05). Age affected several semen quality parameters but did not affect HSP70 gene expression and protein abundance. The HSP70 gene molecule could be a biomarker that determines the fertility of Bali bulls. HSP HSP7 ≤ ( 1 years. . years) computerassisted computer assisted analysis DiffQuick Diff Quick staining FITCPNA FITC PNA qRTPCR, qRTPCR qRT PCR, PCR qRT-PCR immunoassays Indonesianintegrated Indonesian integrated iSIKHNAS. iSIKHNAS (iSIKHNAS) P<0.05 P005 P 0 05 (P<0.05 midpiece P>0.05 (P>0.05 P<0.05. (iSIKHNAS P<0.0 P00 (P<0.0 P>0.0 (P>0.0 P<0. P0 (P<0. P>0. (P>0. P<0 (P<0 P>0 (P>0 P< (P< P> (P> (P

RESUMO O cádmio (Cd) é um poluente ambiental que afeta vários tecidos e órgãos, inclusive o testículo. Muitos estudos demonstraram que a toxicidade do Cd causa comprometimento testicular por meio de estresse oxidativo e ação inflamatória. Portanto, este estudo teve como objetivo demonstrar a toxicidade testicular do Cd e a ação protetora da zeólita contra os efeitos deletérios do cádmio. Ratos machos adultos receberam Cd em uma dose de 30mg/kg/dia por 28 dias consecutivos com ou sem zeólita, que foi administrada em uma dose de 100mg/kg/dia por 28 dias. Foram avaliados o peso dos testículos, os espermatozoides (contagem, motilidade e anormalidades), a testosterona sérica e o hormônio luteinizante (LH), as enzimas testiculares fosfatase ácida (ACP) e fosfatase alcalina (ALP), as citocinas inflamatórias fator de necrose tumoral alfa (TNF-α), interleucina-1beta (IL-1β) e fator nuclear Kappa B (NF-κB) e estresse oxidativo. Aqui, descobrimos que o cádmio causou alterações nas características dos espermatozoides, distúrbios nos hormônios sexuais, declínio nas enzimas testiculares, conteúdo elevado de malondialdeído (MDA), diminuição da glutationa (GSH), aumento do fator nuclear Kappa B (NF-κB) e níveis de citocinas pró-inflamatórias do fator de necrose tumoral alfa (TNF-α) e interleucina-1beta (IL-1β) no homogenato do testículo. Em contraste, a zeólita alterou significativamente esses efeitos deletérios, e o mecanismo em potencial envolveu a regulação negativa do Fator Nuclear Kappa B (NF-κB), do Fator de Necrose Tumoral Alfa (TNF-α) e da interleucina-1beta (IL-1β), restaurando a glutationa (GSH) e reduzindo o malondialdeído (MDA). Além disso, a zeólita foi associada a uma maior taxa de gravidez. Nossos dados sugerem que o estresse oxidativo e a inflamação são responsáveis pela lesão testicular induzida por Cd e que a administração conjunta de zeólita exerce um efeito protetor por meio da via NF-κB /TNF-α/IL-1β. (Cd órgãos testículo inflamatória Portanto 30mgkgdia mgkgdia 30mg kg dia mg 2 100mgkgdia 100mg testículos contagem, contagem (contagem anormalidades, anormalidades , anormalidades) LH, LH (LH) ACP (ACP ALP, ALP (ALP) TNFα, TNFα TNF α interleucina1beta interleucinabeta interleucina 1beta beta IL1β ILβ IL 1β β (IL-1β NFκB NF κB (NF-κB Aqui sexuais MDA, MDA (MDA) GSH, GSH próinflamatórias pró (TNF-α contraste NFκB, IL1β, (GSH MDA. . disso gravidez /TNFα/IL1β. TNFαIL1β TNFαILβ /TNF α/IL 1β. /TNF-α/IL-1β (LH (ALP (MDA /TNFα/IL1β TNFαIL αIL

ABSTRACT Cadmium (Cd) is an environmental pollutant affecting various tissues and organs, including the testis. Many studies demonstrated that Cd toxicity causes testicular impairment through oxidative stress and inflammatory action. Therefore, this study aimed to demonstrate Cd's testicular toxicity and the protective action of zeolite against cadmium's deleterious effects. Adult male rats were given Cd at a dose of 30mg/kg/day for 28 consecutive days with or without zeolite, which was given at a dose of 100mg/kg/day for 28 days. Testis weight, sperm (count, motility, and abnormalities), serum testosterone and luteinizing hormone (LH), testicular enzymes Acid phosphatase (ACP) and Alkaline phosphatase(ALP), inflammatory cytokines Tumor Necrosis Factor Alpha (TNF-α) , interleukin-1beta (IL-1β) , and Nuclear Factor Kappa B (NF-κB) and oxidative stress were evaluated. Herein, we found that cadmium caused alterations in sperm characteristics, sex hormone disturbance, decline in testicular enzymes, elevated malondialdehyde (MDA) contents, decreased glutathione (GSH), increased Nuclear Factor Kappa B (NF-κB) and pro-inflammatory cytokines Tumor Necrosis Factor Alpha (TNF-α) and interleukin-1beta (IL-1β) levels in testis homogenate. In contrast, zeolite significantly amended these deleterious effects, and the potential mechanism involved the downregulation of Nuclear Factor Kappa B (NF-κB), Tumor Necrosis Factor Alpha (TNF-α) and interleukin-1beta (IL-1β), restoring glutathione (GSH) and reducing malondialdehyde (MDA). Also, zeolite was associated with an increased rate of pregnancy. Our data suggested that oxidative stress and inflammation are responsible for Cd-induced testicular injury and co-administration of zeolite exerts a protective effect via NF-κB /TNF-α/IL-1β pathway. (Cd organs Therefore Cds s cadmiums effects 30mgkgday mgkgday 30mg kg day mg 2 100mgkgday 100mg weight count, count (count motility abnormalities, abnormalities abnormalities) LH, LH (LH) ACP (ACP phosphataseALP, phosphataseALP ALP phosphatase(ALP) TNFα TNF α (TNF-α interleukin1beta interleukinbeta interleukin 1beta beta IL1β ILβ IL 1β β (IL-1β NFκB NF κB (NF-κB evaluated Herein characteristics disturbance MDA (MDA contents GSH, GSH proinflammatory pro homogenate contrast NFκB, IL1β, (GSH MDA. . Also pregnancy Cdinduced induced coadministration co administration /TNFα/IL1β TNFαIL1β TNFαILβ /TNF α/IL pathway (LH phosphatase(ALP TNFαIL αIL

Abstract The current investigation aimed to explore the effects of Myrciaria dubia liquid extract (MDLE) as the primary component of an extender for breeder rooster semen over different periods at room temperature. Fifteen breeder roosters (40 weeks of age, average body weight of 2.05±0.12) with confirmed fertility were used. Employing a factorial design (3x4), the treatments consisted of semen in natura and two semen extenders (an experimental based on MDLE and a commercial) subjected to four periods at room temperature post-collection (5, 10, 15 and 20 minutes) with four replicates (tubes) each. All variables evaluated in this study yielding significant results (p<0.05). Analyzed individually, the experimental extender based on MDLE exhibited a linear reduction (p<0.05) in motility and vigor results, while it caused an increase in pH values and percentages of sperm defects evaluated. When compared with semen in natura and commercial extender, the efficiency of MDLE as a semen extender was inferior to that observed with the commercial extender and similar to the results observed with semen in natura. Nonetheless, the experimental extender based on MDLE yielded satisfactory results for up to 15 minutes of storage time. In conclusion, MDLE can be considered as an alternative for composing a roosters’ semen extender, maintaining sperm characteristics within acceptable limits for up to 15 minutes at room temperature. However, this experimental extender demonstrated lower efficiency than the commercial extender in maintaining the sperm quality at room temperature across all periods tested. (MDLE 40 (4 age 2.05±0.12 205012 2 05 0 12 used 3x4, 3x4 x , (3x4) postcollection post collection 5, 5 (5 10 1 tubes (tubes each p<0.05. p005 p p<0.05 . individually (p<0.05 Nonetheless time conclusion However tested 4 ( 2.05±0.1 20501 3x (3x4 p00 p<0.0 (p<0.0 2.05±0. 2050 (3x p0 p<0. (p<0. 2.05±0 205 p<0 (p<0 2.05± p< (p< 2.05 (p 2.0 2.

The objective of this study was to assess semen characteristics of bucks kept under communal management systems in the hard-veld, southern part of Botswana. In early autumn, semen was collected from 22 bucks ranging from < 2 to >5 years old across 20 randomly-selected communal farms using an electro-ejaculator. The buck type, age, live weight, and scrotal circumference were recorded prior to semen collection. Semen characteristics assessed were semen colour, ejaculated volume, sperm cell concentration, sperm motility, as well as morphology. The data were examined using analysis of variance with a general linear model in SAS. The common buck breed kept by communal farmers was a non-descript (mixed) breed (86%) and the Boer goat (14%) was in minority. The most predominant semen colour was creamy. The breed did not influence live weight, scrotal circumference, and semen characteristics. The younger bucks (<2 years) had substantially lower live weight, scrotal circumference, and ejaculated semen volume than the older bucks. Semen characteristics did not differ substantially across the ages except for the sperm concentration, which was lower in bucks <2 years compared to >5 years. Live weight was strongly correlated (r = 0.81) to scrotal circumference and moderately correlated (r = 0.60) to the ejaculated semen volume. The study provides an insight into semen quality and prediction of fertility for bucks kept under a communal management system.

Resumo A qualidade espermática é parâmetro fundamental para a reprodução eficaz de peixes em cativeiro e para o desenvolvimento de técnicas reprodutivas, como a criopreservação de sêmen. Este estudo teve como objetivo determinar a composição do plasma seminal de Pseudoplatystoma reticulatum e analisar as relações entre os componentes do plasma seminal e as características espermáticas. Nove machos foram induzidos à espermiação com extrato de hipófise de carpa no período reprodutivo da espécie (novembro e dezembro/2019). As características do sêmen foram avaliadas: motilidade espermática subjetiva, motilidade, duração, volume espermático liberado, concentração espermática, pH, osmolalidade e composição do plasma seminal, incluindo níveis de cálcio, cloreto, sódio, magnésio, potássio, glicose, frutosamina, triglicerídeos e proteína total. Para determinar a relação entre os componentes do plasma seminal e os parâmetros de motilidade espermática, foi realizada uma análise de componentes principais (PCA). O plasma seminal de P. reticulatum é composto principalmente pelo íon Na+ e componentes orgânicos como proteínas e glicose. Através da PCA, observou-se que a motilidade espermática apresentou forte correlação positiva com tempo de motilidade, concentração espermática e proteína total e correlação negativa com osmolalidade e frutosamina. reprodutivas espermáticas novembro dezembro/2019. dezembro2019 dezembro dezembro/2019 . 2019 dezembro/2019) avaliadas subjetiva duração liberado pH cálcio cloreto sódio magnésio potássio glicose frutosamina PCA. PCA (PCA) P Na observouse observou se dezembro201 dezembro/201 201 (PCA dezembro20 dezembro/20 20 dezembro2 dezembro/2 2 dezembro/

Abstract Sperm quality is a fundamental parameter for the effective reproduction of fish in captivity and the development of reproductive techniques, such as semen cryopreservation. This study aimed to determine the composition of the seminal plasma of Pseudoplatystoma reticulatum and analyze the relationships between plasma components and sperm characteristics. Nine males were induced to spermiation with carp pituitary extract in the reproductive period of the species (November and December/2019). Semen characteristics were evaluated: subjective sperm motility, motility, duration, released sperm volume, sperm concentration, pH, osmolality, and seminal plasma composition, including levels of calcium, chloride, sodium, magnesium, potassium, glucose, fructosamine, triglycerides, and total protein. To determine the relationship between seminal plasma components and sperm motility parameters, a principal component analysis (PCA) was performed. The seminal plasma of P. reticulatum is composed mainly of the Na+ ion and organic components such as protein and glucose. Through PCA, it was observed that sperm motility had a strong positive correlation with motility time, sperm concentration, and total protein and a negative correlation with osmolality and fructosamine. techniques cryopreservation November December/2019. December2019 December December/2019 . 2019 December/2019) evaluated duration volume concentration pH calcium chloride sodium magnesium potassium glucose fructosamine triglycerides parameters PCA (PCA performed P Na time December201 December/201 201 December20 December/20 20 December2 December/2 2 December/

ABSTRACT The Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae), is one of the most harmful species to world horticulture, thus requiring suppression or eradication programs. The sterile insect technique is widely used for autocidal control of nuisance insects because it neither releases exotic agents into the environment nor introduces new genetic material into existing populations. In 2005, the Moscamed Brasil biofactory imported the tsl Vienna 8 C. capitata strain from the International Atomic Energy Agency (IAEA); this action required genetic testing of the strain’s biological characteristics. The objective of this study was to discriminate males of wild populations from tsl Vienna 8 using molecular markers, and to conclude which male line dominated in a remating condition. Four crosses, each using 100 males and 100 females, were performed: wild female × wild male; wild female × tsl Vienna 8 male; wild female × wild male, followed by remating with tsl Vienna 8 males after 48 h, and wild female × tsl Vienna 8 male, followed by remating with wild males after 48 h. The results showed that the tsl Vienna 8 strain is compatible with wild females from the São Francisco Valley region and that these males can successfully transfer their sperm to the female spermathecae. Regarding remating, the sperm of the second male dominated over that of the first one. Based on these findings, the sterile insect technique success may be reduced, since the progeny of a female that remates with a wild male could be viable. fly Diptera (Diptera Tephritidae, Tephritidae , Tephritidae) horticulture programs 2005 C IAEA (IAEA) strains s characteristics markers condition crosses 10 performed 4 h spermathecae findings reduced viable 200 (IAEA 1 20 2

RESUMO A mosca-das-frutas do Mediterrâneo, Ceratitis capitata (Diptera: Tephritidae), é uma das espécies mais prejudiciais à horticultura mundial, portanto requer programas de supressão ou erradicação. A técnica do inseto estéril é amplamente utilizada para o controle autocida de insetos nocivos, pois não libera agentes exóticos no meio ambiente nem introduz novos materiais genéticos nas populações existentes. Em 2005, a biofábrica Moscamed Brasil importou a cepa Vienna 8 C. capitata da Agência Internacional de Energia Atômica (IAEA); essa ação exigiu testes genéticos das características biológicas da linhagem. O objetivo deste estudo foi discriminar machos de populações silvestres de tsl Viena 8 usando marcadores moleculares, e inferir sobre qual linhagem masculina era dominante em condição de recópula. Quatro cruzamentos, cada um com 100 machos e 100 fêmeas, foram realizados: fêmea selvagem × macho selvagem; fêmea selvagem × macho tsl Viena 8; fêmea selvagem × macho selvagem, seguido por recópula com machos tsl Vienna 8 após 48 h, e fêmea selvagem × machos tsl Vienna 8, seguido por recópula com machos selvagens após 48 h. Os resultados mostraram que a cepa tsl Vienna 8 é compatível com fêmeas selvagens da região do Vale do São Francisco e que esses machos podem transferir com sucesso seus espermatozóides para a espermateca das fêmeas. Quanto à recópula, o esperma do segundo macho precedeu o primeiro. Com base nesses achados, o sucesso da técnica de insetos estéril pode ser reduzido, já que a progênie de uma fêmea que recopulou com um macho selvagem pode ser viável. moscadasfrutas mosca frutas Mediterrâneo Diptera (Diptera Tephritidae, Tephritidae , Tephritidae) mundial erradicação nocivos existentes 2005 C IAEA (IAEA) moleculares cruzamentos 10 realizados 4 h primeiro achados reduzido viável 200 (IAEA 1 20 2

Resumen El toro es un factor de importancia genética y económica en un hato ganadero, ya que de este dependerán las próximas generaciones; por lo tanto, es esencial la evaluación de la calidad seminal y la criopreservación de su material genético. El objetivo del estudio fue evaluar el efecto de dos temperaturas de dilución sobre la calidad de semen de ganado cebuino del Trópico Peruano. Dos toros Gyr fueron colectados semanalmente mediante electroeyaculación. Las muestras se colocaron en baño maría a 34°C y se analizaron las características macroscópicas y microscópicas del semen. Solo los eyaculados con >60% de motilidad y <20% de anormalidades fueron procesados, separándolos en dos tratamientos. El T1 consistió en predilución 1:1 a 34°C, luego de 30 minutos una dilución final agregando el dilutor a 34°C. El T2 consistió en predilución 1:1 a 34°C y a los 30 minutos de estabilización se llevó a refrigeración conjuntamente con el dilutor, para luego realizar la dilución final a 5°C. La calidad seminal fue analizada luego de 16 horas de refrigeración. El porcentaje de vitalidad en T1 (80,89%) fue mayor que en T2 (71,22%) (p<0,05), la reacción positiva a la prueba hipoosmótica fue mayor en T1 (71,11%) que en T2 (60%) (p<0,05). La integridad acrosomal y los rasgos morfométricos no variaron según tratamientos. En conclusión, la dilución inicial y final a 34°C en toros Gyr influye positivamente sobre la calidad seminal antes de la congelación de pajillas de semen.

Abstract The bull is a factor of genetic and economic importance in cattle breed, since the next generations will depend on it; therefore, the evaluation of the seminal quality and the cryopreservation of its genetic material are essential. The aim of the study was to evaluate the effect of two dilution temperatures on the quality of semen from zebuine cattle from the Peruvian Tropics. The samples were placed in a water bath at 34°C and the macroscopic and microscopic characteristics of semen were analyzed. Only the ejaculates with >60% motility and <20% abnormalities were processed, separating them into two treatments. T1 consisted of a 1:1 predilution at 34°C, followed by a final dilution of 30 minutes by adding the dilutor at 34°C. T2 consisted of a 1:1 predilution at 34°C and after 30 minutes of stabilization, refrigeration was carried out together with the dilutor, in order to carry out the final dilution at 5°C. The seminal quality was analyzed after 16 hours of refrigeration. The percentage of vitality in T1 (80.89%) was greater than in T2 (71.22%) (p<0.05), the positive reaction to the hypoosmotic problem was greater in T1 (71.11%) than in T2 (60%) (p<0.05). Acrosomal integrity and morphometric features did not vary according to treatments. In conclusion, the initial and final dilution at 34°C in Gyr bulls positively influences the semen quality before the freezing of semen packets.

Abstract There is a paucity of information with respect to group-training for artificial vagina and its influence on semen characteristics and sexual behavior of young untrained rams. A total of 18 healthy Najdi rams (with an initial body weight of 40-45 Kg and 7-8 month-old) were consequently used herein to test the usefulness of group-training for artificial vagina-mediated semen collection during the breeding season. Rams were randomly segregated into three groups (n = 6 rams per protocol), and the whole experiment was lasted for 10 weeks. The 1st group was subjected to a training protocol where one untrained ram was placed for 20 min with a teaser ewe, while the 2nd group were subjected to a protocol where one untrained ram was placed for 20 min with one trained ram and a teaser ewe, whereas the 3rd group were subjected to a protocol where three untrained rams were placed for 20 min with one trained ram and a teaser ewe. The obtained results clearly (P < 0.05) showed that training young rams in group has increased their sperm concentration and sexual stimulation, shortened the period of their training time, and descriptively had a complete training efficiency. The sexual stimulation of young untrained rams was intensified by the competition between rams in the co-presence of a trained ram. Collectively, these data may suggest that group-training of rams at puberty is a better protocol for AV-mediated semen collection compared to individual training. Some shortcomings were noted herein, but research dealing with this subject may very well improve the reproductive performance of young untrained rams. grouptraining 1 4045 40 45 40-4 78 7 8 7- monthold month old month-old vaginamediated mediated season n protocol, , protocol) weeks st 2 ewe nd rd P 0.05 005 0 05 time efficiency copresence co presence Collectively AVmediated AV 404 4 40- 0.0 00 0.

ABSTRACT. This study evaluated the effects of chromium picolinate (CrPic) with or without vitamin C on the reproductive potentials of cocks raised under high ambient temperature. Four diets contained CrPic at 0.00 to 1.20 mg kg-1 without vitamin C and another four diets contained CrPic with 200 mg vitamin C each. A total of 192 White Leghorn and Noiler cocks (96 cocks each) were randomly assigned to the 8 treatments and replicated 4 times with 6 cocks per replicate in a 2 x2 x 4 factorial experiment. At the end of the sixteen weeks study, twelve birds per treatment were randomly sacrificed. The testes were carefully sampled, weighed and processed for estimation of daily sperm production using the homogenate method. The results revealed that the interactions of 0.40 mg CrPic kg-1 with or without vitamin C significantly (p < 0.05) influenced the gonadal and extra-gonadal weights, semen characteristics among the cocks while the daily sperm production and reproductive hormones were not negatively affected. This study, therefore, concluded that 0.40 mg CrPic kg-1 diet with vitamin C could significantly reduce the effects of heat stress on cocks raised under high ambient temperature without negatively impacting on their normal reproductive functions. ABSTRACT (CrPic 000 0 00 0.0 120 1 20 1.2 kg1 kg kg- each 19 96 (9 experiment sacrificed sampled method 040 40 0.4 p 0.05 005 05 extragonadal extra weights affected therefore functions 0. 12 1. 9 ( 04

ABSTRACT The objective of this study was to evaluate the effect of the seasons (Summer and Autumn), on live weight, body condition, mass motility, percentage of live spermatozoa, and sperm-cell concentration of Creole roosters (Gallus domesticus) from Mexico. Semen from 35-week-old Creole roosters was collected weekly during 10 weeks in Summer and Autumn, through the dorso-abdominal massage technique. Roosters were individually kept under a constant photoperiod (16 hours light:8 hours dark). The average live weight was 4.5% higher (p<0.05) in Autumn (2.78 kg) than in Summer (2.66 kg), therefore this variable increased with age (r = 0.85, p<0.05). Category 2 of body condition occurred (p<0.05) with higher probability than the others (0, 1 and 3), being practically the same (p>0.05) in Autumn (99.96%) and in Summer (99.81%). On average (and in weeks 1 and 3-10), the percentage of live spermatozoa was higher in Summer than in Autumn. Accordingly, the percentage of live spermatozoa decreased with age (r = -0.82, p<0.05). However, on average, sperm-cell concentration did not change between seasons (p>0.05). In conclusion, Mexican Creole roosters showed higher percentage of live spermatozoa in Summer than in Autumn. Therefore, it is advisable to select these animals of about 2.7 kg and reproduce them in Summer. , Autumn) motility spermcell sperm cell Gallus domesticus Mexico 35weekold weekold 35 week old dorsoabdominal dorso abdominal technique 16 (1 light8 light 8 light: dark. dark . dark) 45 4 5 4.5 p<0.05 p005 p 0 05 (p<0.05 2.78 278 78 (2.7 2.66 266 66 (2.6 kg, r 085 85 0.85 p<0.05. p<0.05) 0, (0 3, 3 3) p>0.05 (p>0.05 99.96% 9996 99 96 (99.96% 99.81%. 9981 99.81% 81 (99.81%) 310, 310 3-10) Accordingly 0.82, 082 82 -0.82 However p>0.05. conclusion Therefore 27 7 2. ( 4. p<0.0 p00 (p<0.0 (2. 2.6 26 6 08 0.8 p>0.0 (p>0.0 99.96 999 9 (99.96 998 99.81 (99.81% 31 3-10 0.82 -0.8 p<0. p0 (p<0. (2 0. p>0. (p>0. 99.9 (99.9 99.8 (99.81 3-1 -0. p<0 (p<0 p>0 (p>0 99. (99. (99.8 3- -0 p< (p< p> (p> (99 - (p (9

ABSTRACT The objective of this study was to evaluate characteristics of the testicular parenchyma and vascular parameters of the pampiniform plexus obtained by ultrasound, semen quality parameters, and sperm freezability in Nellore bulls classified based on residual feed intake (RFI). Twenty-seven bulls (21.82±0.88 months of age) evaluated for feed efficiency were sampled for the study, including 15 with low RFI (−0.592±0.09 kg dry matter/day) and 12 with high RFI (0.792±0.10 kg dry matter/day). In ultrasound and Doppler assessment, the most efficient animals (low RFI) showed higher pulsatility and resistive indexes, as well as a tendency towards greater heterogeneity of the testicular parenchyma (0.625±0.032 vs. 0.508±0.032, 1.012±0.072 vs. 0.802±0.072, and 12.9±0.96 vs. 10.2±0.96, respectively, for low vs. high RFI). However, these animals tended to have lower peak diastolic velocity (5.19±0.50 for low RFI vs. 6.54±0.50 for high RFI). Analysis of fresh semen showed a lower percentage of minor defects in low RFI animals (2.67±1.19%) compared with high RFI animals (8.10±1.19%), without differences in the other parameters in fresh or thawed semen and after thermoresistance testing. Evaluation of flow cytometry parameters showed a higher quality of mitochondrial respiration in semen samples of low RFI animals (22.04±2.50%) compared with high RFI animals (12.29±2.71%). Therefore, although RFI exerts an effect on the Doppler parameters of the pampiniform plexus, it is not sufficient to affect the quality of fresh or thawed semen. RFI. . (RFI) Twentyseven Twenty seven 21.82±0.88 2182088 21 82 0 88 (21.82±0.8 age 1 −0.592±0.09 0592009 592 09 (−0.592±0.0 matter/day matterday matter day 0.792±0.10 0792010 792 10 (0.792±0.1 matter/day. assessment indexes 0.625±0.032 06250032 625 032 (0.625±0.03 vs 05080032 508 0.508±0.032 10120072 012 072 1.012±0.07 08020072 802 0.802±0.072 129096 9 96 12.9±0.9 102096 2 10.2±0.96 respectively However 5.19±0.50 519050 5 19 50 (5.19±0.5 654050 6 54 6.54±0.5 2.67±1.19% 267119 67 (2.67±1.19% 8.10±1.19%, 810119 8.10±1.19% , 8 (8.10±1.19%) testing 22.04±2.50% 2204250 22 04 (22.04±2.50% 12.29±2.71%. 1229271 12.29±2.71% 29 71 (12.29±2.71%) Therefore (RFI 21.82±0.8 218208 (21.82±0. −0.592±0.0 059200 59 (−0.592±0. 0.792±0.1 079201 79 (0.792±0. 0.625±0.03 0625003 62 03 (0.625±0.0 0508003 0.508±0.03 1012007 01 07 1.012±0.0 0802007 80 0.802±0.07 12909 12.9±0. 10209 10.2±0.9 5.19±0.5 51905 (5.19±0. 65405 6.54±0. 2.67±1.19 26711 (2.67±1.19 81011 8.10±1.19 (8.10±1.19% 22.04±2.50 220425 (22.04±2.50 122927 12.29±2.71 7 (12.29±2.71% 21.82±0. 21820 (21.82±0 −0.592±0. 05920 (−0.592±0 0.792±0. 07920 (0.792±0 0.625±0.0 062500 (0.625±0. 050800 0.508±0.0 101200 1.012±0. 080200 0.802±0.0 1290 12.9±0 1020 10.2±0. 5.19±0. 5190 (5.19±0 6540 6.54±0 2.67±1.1 2671 (2.67±1.1 8101 8.10±1.1 (8.10±1.19 22.04±2.5 22042 (22.04±2.5 12292 12.29±2.7 (12.29±2.71 21.82±0 2182 (21.82± −0.592±0 0592 (−0.592± 0.792±0 0792 (0.792± 0.625±0. 06250 (0.625±0 05080 0.508±0. 10120 1.012±0 08020 0.802±0. 129 12.9± 102 10.2±0 5.19±0 519 (5.19± 654 6.54± 2.67±1. 267 (2.67±1. 810 8.10±1. (8.10±1.1 22.04±2. 2204 (22.04±2. 1229 12.29±2. (12.29±2.7 21.82± 218 (21.82 −0.592± 059 (−0.592 0.792± 079 (0.792 0.625±0 0625 (0.625± 0508 0.508±0 1012 1.012± 0802 0.802±0 12.9 10.2± 5.19± 51 (5.19 65 6.54 2.67±1 26 (2.67±1 81 8.10±1 (8.10±1. 22.04±2 220 (22.04±2 122 12.29±2 (12.29±2. 21.82 (21.8 −0.592 05 (−0.59 0.792 (0.79 0.625± 062 (0.625 050 0.508± 101 1.012 080 0.802± 12. 10.2 5.19 (5.1 6.5 2.67± (2.67± 8.10± (8.10±1 22.04± (22.04± 12.29± (12.29±2 21.8 (21. −0.59 (−0.5 0.79 (0.7 0.625 06 (0.62 0.508 1.01 08 0.802 10. 5.1 (5. 6. 2.67 (2.67 8.10 (8.10± 22.04 (22.04 12.29 (12.29± 21. (21 −0.5 (−0. 0.7 (0. 0.62 (0.6 0.50 1.0 0.80 5. (5 2.6 (2.6 8.1 (8.10 22.0 (22.0 12.2 (12.29 (2 −0. (−0 0. (0 0.6 0.5 1. 0.8 ( 2. (2. 8. (8.1 22. (22. (12.2 −0 (− (8. (22 (12. − (8 (12 (1

Abstract The proposed study was to determine if the silver nanoparticles can be used as potential antimicrobial agents and can replace the use of conventional antibiotics in semen without affecting the motility and fertility of semen. The silver nanoparticles prepared by chemical reduction method were confirmed by determination of the wavelength of surface plasmon resonance peak and further characterized using Zetasizer by determining their size, polydispersity index, and zeta potential. The nanoparticles were assessed for antibacterial activity and their concentration was optimized for use in semen extender for cryopreservation. Cryopreserved semen was further evaluated for seminal parameters, antioxidant parameter, and microbial load. Prepared silver NPs showed a plasmon resonance peak at 417 nm wavelength. NPs were found to possess antibacterial activity and were supplemented in semen extender @ 125 and 250 µg/ml for semen cryopreservation. There was a significant increase in pre and post-freezing motility and other seminal parameters. The microbial load of frozen-thawed semen of control and supplemented groups were well within the permissible limits. Lipid peroxidation levels were reduced in NPs supplemented groups, and reactive oxygen species (ROS) levels were significantly reduced in semen supplemented with 125 µg/ml NPs. Thus it can be conclude that silver NPs can be successfully used as a substitute for antibiotics in cattle bull semen cryopreservation with good antimicrobial activity and no adverse effects on sperm characteristics. size index parameters parameter 41 12 25 µgml µg ml postfreezing post freezing frozenthawed frozen thawed limits ROS (ROS characteristics 4 1 2

Abstract Up to now, the definitive conclusion of the positive effects of rapid transient thawing at higher temperatures for shorter durations has not been obtained yet and is still under discussion due to some contradictory findings and limited assessment of post-thawed parameters. The purpose of the current study was to evaluate the effectiveness of rapid thawing in water at 70 °C by using various post-thawed parameters of frozen bull spermatozoa. Experiment 1, monitoring the change of temperature inside frozen bull straw thawed in water at different temperatures. Experiment 2, evaluation of various post-thawed characteristics of frozen bull spermatozoa thawed in water at different temperatures by using a computer-assisted sperm analysis, flow cytometry and immunocytochemistry. The time it took for the temperature inside the straw to warm up to 15 °C was nearly twice as faster when the straw was thawed in 70 °C water compared with 39 °C. Although there were differences among bulls, viability, motility, and mitochondrial membrane potential of spermatozoa thawed at 70 °C for 8 seconds and stabilized at 39 °C for 52 seconds were significantly higher than those of controls (thawed at 39 °C for 60 seconds) at 0 and 3 h after thawing. Just after thawing, however, there were no differences in acrosome integrity and distribution of phospholipase C zeta1, whereas mitochondrial reactive oxygen species production was significantly lower in spermatozoa thawed at 70 °C. From these results, we conclude that rapid thawing at 70 °C and then stabilization at 39 °C significantly improves viability, motility and mitochondrial health of bull spermatozoa rather than conventional thawing at 39 °C. The beneficial effect of rapid transient thawing could be due to shorter exposure to temperatures outside the physiological range, consequently maintaining mitochondrial health. now postthawed post 7 1 2 computerassisted computer assisted analysis immunocytochemistry bulls viability 5 6 however zeta1 zeta results range

Abstract Background Infertility is caused by heterogeneous risks, but most of them are unexplained. The sperm DNA Fragmentation Index (DFI) was increasingly acknowledged as a parameter for the evaluation of male infertility. This study aimed to investigate the association between sperm DFI and laboratory and clinical outcomes in a population with unexplained infertility. Methods The clinical data of an infertile population was collected for the selection of reproductive patients with unexplained infertility. The authors classified the patients with normal sperm parameters in a control group (DFI < 25%) and an observation group (DFI ≥ 25%) and compared the difference in basal characteristics, laboratory, and clinical outcomes between the two groups. The authors conducted a correlation analysis to examine the relationship between DFI and the number of D3 good-quality embryos, as well as the clinical pregnancy rate and live birth rate. A total of 176 cases were enrolled in the retrospective study. Results The observation group (n = 88) showed advanced male age, lower sperm concentration, progressive motility, and morphology assessment than the control group. In addition, lower No. of D3 good-quality embryos, clinical pregnancy rate, and the live birth rate were shown in the observation group. A negative correlation between the DFI and No. of D3 good-quality embryos (rs = -0.347, p < 0.001) or live birth rate (rs = -0.185, p = 0.028) was shown. Conclusions Sperm DFI was a good indicator for the prediction of D3 good-quality embryos in unexplained infertility couples, but it did not provide sufficient information regarding clinical pregnancy outcome but live pregnancy outcome. risks 25% 25 characteristics groups D goodquality quality 17 n 88 age concentration motility addition No rs 0.347, 0347 0 347 -0.347 0.001 0001 001 0.185, 0185 185 -0.185 0.028 0028 028 couples 2 1 8 0.347 034 34 -0.34 0.00 000 00 0.185 018 18 -0.18 0.02 002 02 0.34 03 3 -0.3 0.0 0.18 01 -0.1 0.3 -0. 0. 0.1 -0 -

Abstract The objective of this study was to obtain data on the testicular biometry of the white-lipped peccary (Tayassu pecari) and to verify if there is a correlation between the individual’s biometric data with their dominance rank. Data were collected from 16 adult males, aged between two and seven years, who made up the same group. By the agonistic interactions analyses we calculated the linearity indexes (h') of the dominance hierarchy and dominance rank. At the end of the behavioral observations, males were captured to collect biometric data from the testis. White-lipped peccaries showed testicles located in a perineal position, inclined cranio-ventrally with oval shape, flattened laterally and with tenso-elastic consistency (2.54±0.07). There was variance in the means (± standard deviation) of testis length (5.88±1.05cm), width (4.24 ± 0.98cm), height (4.44±0.86cm), and total scrotal width (8.78±17.05). The linear dominance hierarchy described the social structure of the males of this group (h'>0.9), with probability of linearity in the hierarchy greater than chance (P = 0.02). There was a correlation between dominance rank with the body mass, length and volume of the testicles. As in general the dimensions of the testicles are directly correlated with sperm production and also testosterone, this characteristic favors the reproductive performance of dominant white-lips males. Therefore, the characteristics of testicular biometry of white-lips should be considered for the selection of individuals more likely to reproduce in captivity favoring the conservation of this vulnerable species. whitelipped white lipped Tayassu pecari individual s 1 years h (h' observations Whitelipped White position cranioventrally cranio ventrally shape tensoelastic tenso elastic 2.54±0.07. 254007 2.54±0.07 . 2 54 0 07 (2.54±0.07) ( deviation 5.88±1.05cm, 588105cm cm 5.88±1.05cm , 5 88 05cm (5.88±1.05cm) 4.24 424 4 24 (4.2 0.98cm, 098cm 0.98cm 98cm 0.98cm) 4.44±0.86cm, 444086cm 4.44±0.86cm 44 86cm (4.44±0.86cm) 8.78±17.05. 8781705 8.78±17.05 8 78 17 05 (8.78±17.05) h>0.9, h09 >0.9 9 (h'>0.9) P 0.02. 002 0.02 02 0.02) mass testosterone whitelips lips Therefore species (h 25400 2.54±0.0 (2.54±0.07 (5.88±1.05cm 4.2 42 (4. (4.44±0.86cm 878170 8.78±17.0 7 (8.78±17.05 h>0.9 h0 09 >0. (h'>0.9 00 0.0 2540 2.54±0. (2.54±0.0 4. (4 87817 8.78±17. (8.78±17.0 h>0. >0 (h'>0. 0. 254 2.54±0 (2.54±0. 8781 8.78±17 (8.78±17. h>0 > (h'>0 25 2.54± (2.54±0 878 8.78±1 (8.78±17 h> (h'> 2.54 (2.54± 87 8.78± (8.78±1 2.5 (2.54 8.78 (8.78± 2. (2.5 8.7 (8.78 (2. 8. (8.7 (2 (8. (8

Resumo O objetivo neste estudo foi obter dados sobre a biometria testicular do queixada (Tayassu pecari) e verificar se há correlação entre os dados biométricos do indivíduo com seu posto de dominância. Os dados foram coletados de 16 queixadas adultos, com idades entre dois e sete anos, que compunham o mesmo grupo. Por meio de análises das interações agonísticas foram calculados os índices de linearidade (h') da hierarquia de dominância e determinado o rank dos indivíduos. Ao final das observações comportamentais, os machos foram capturados para coleta de dados biométricos do testículo. Os queixadas apresentaram testículos localizados em posição perineal, inclinados cranioventralmente com formato oval, achatados láterolateralmente e com consistência tensoelástica (2,54±0,07). Houve variação nas médias (± desvio padrão) do comprimento testicular (5,88±1,05cm), largura (4,24±0,98cm), altura (4,44±0,86cm) e largura escrotal total (8,78±17,05). A hierarquia de dominância linear descreveu a estrutura social dos machos desse grupo (h'>0,9), com probabilidade de linearidade na hierarquia maior que o acaso (P = 0,02). Houve correlação entre a classificação de dominância com a massa corporal, comprimento e volume dos testículos. Como em geral as dimensões dos testículos estão diretamente correlacionadas com a produção de esperma e também de testosterona, essa característica favorece o desempenho reprodutivo de machos de queixadas brancos dominantes. Portanto, as características da biometria testicular de queixadas devem ser consideradas para a seleção de indivíduos com maior probabilidade de reprodução em cativeiro favorecendo a conservação desta espécie vulnerável. Tayassu pecari 1 adultos anos h (h' comportamentais testículo perineal oval 2,54±0,07. 254007 2,54±0,07 . 2 54 0 07 (2,54±0,07) ± ( padrão 5,88±1,05cm, 588105cm cm 5,88±1,05cm , 5 88 05cm (5,88±1,05cm) 4,24±0,98cm, 424098cm 4,24±0,98cm 4 24 98cm (4,24±0,98cm) 4,44±0,86cm 444086cm 44 86cm (4,44±0,86cm 8,78±17,05. 8781705 8,78±17,05 8 78 17 05 (8,78±17,05) h>0,9, h09 >0,9 9 (h'>0,9) P 0,02. 002 0,02 02 0,02) corporal testosterona dominantes Portanto vulnerável (h 25400 2,54±0,0 (2,54±0,07 (5,88±1,05cm (4,24±0,98cm 878170 8,78±17,0 7 (8,78±17,05 h>0,9 h0 09 >0, (h'>0,9 00 0,0 2540 2,54±0, (2,54±0,0 87817 8,78±17, (8,78±17,0 h>0, >0 (h'>0, 0, 254 2,54±0 (2,54±0, 8781 8,78±17 (8,78±17, h>0 > (h'>0 25 2,54± (2,54±0 878 8,78±1 (8,78±17 h> (h'> 2,54 (2,54± 87 8,78± (8,78±1 2,5 (2,54 8,78 (8,78± 2, (2,5 8,7 (8,78 (2, 8, (8,7 (2 (8, (8