Effect of GnRH administration on pregnancy-associated glycoproteins in dairy sheep with different reproductive status

Background: Measurement of pregnancy-associated glycoproteins (PAGs) by radioimmunoassay or enzyme-linked immunosorbent assay has been commonly used for early pregnancy diagnosis in ruminants. However, an accurately pregnancy detection depends on test antibody, breed and number of embryos. Only few studies have conducted to detect or predict animals at risk of late embrionic mortality (LEM) and to use hormonal interventions for embryo losses reducing, but this area is still open in sheep. The current study aimed to evaluate the effect of gonadotropin releasing hormone (GnRH) administration on Day 4 after artificial insemination on the pregnancy-associated glycoproteins in dairy sheep with different reproductive status in relation to pregnancy and late embrionic mortality detection. Materials, Methods & Results: Sixty-five East Friesian sheep were divided in 2 groups - I (Control group, n = 35) and II (GnRH group, n = 30) and sublected to estrus synchronization and artificial insemination (AI). Group I was not treated and Group II received 50 µg GnRH on Day 4 after AI. PAGs in blood serum were measured by Alertys Ruminant Pregnancy test on Days 4, 12, 20, 25 and 35 after AI and ultrasound pregnancy test was conducted on Days 20, 25, 35 and 60. Reproductive status (non-pregnant, pregnant and animals with LEM) was determined by ultrasound and the results between different groups were compared. The PAGs mean values according to reproductive status and Day after AI were analysed. Accuracy, sensitivity and specificity of the PAGs test for pregnancy diagnosis on Day 35 were calculated. On Day 20 after AI the pregnat sheep (83.3%) in group I tended to be higher than those (77.1%) in group II with decrease of 25.7% and 20% for the same groups on Day 25. On day 25 LEM was recorded in 33.3% and 24% in group I and II, respectively. The ultrasound exams on Day 60 confirmed the results from Day 25 after AI. A total value of non-pegnant, pregnant animals and LEM was 20%, 56.9% and 28.8%, respectively. The mean values of PAGs in animals with the same reproductive status in group I and II no differed statistically between Days 4 and 35 after AI. On Day 25 the PAGs values in LEM groups (0.126 ± 0.072 and 0.179 ± 0.029) were higher than those (0.062 ± 0.038 and 0.083 ± 0.023) in the non-pregnant groups, but no significan difference was deternined. On Day 35 after AI the mean values of PAGs (0.414 ± 0.125 and 0.421 ± 0.121) for the pregnant groups were significantly (P < 0.05) higher than those (0.078 ± 0.053 and 0.093 ± 0.034) for the non-pregnant groups. The values of PAGs in LEM groups on Day 25 (0.099 ± 0.062 and 0.113 ± 0.058) were decreased and close to the mean values in non-pregnant sheep. A significant effect of the day after AI on the PAGs values was evidenced in pregnant animals only (control r = 0.97 and GnRH r = 0.98; P < 0.05). The PAGs started to rise rapidly between Days 25 and 35 after AI. On Day 35 the accuracy (98%), the sensitivity (97.3%) and the specificity (100%) of the PAG test for pregnancy diagnosis were similar to the values (100%) for ultrasound method. Discussion: The gonadotropin treatment on Day 4 after AI tended to improve the reproductive performance in the sheep, but no significant effect of GnRH on the PAGs values in the animals with different reproductive status was found. The PAGs profiles in non-pregnant and LEM sheep were close and distinguishing between non-pregnant and LEM ewes on Day 35 was impossible. The PAGs values in pregnant sheep showed significantly (P < 0.05) increase between Days 25 and 35 after AI with higher mean values in pregnant than non-pregnant groups on Day 35 (P < 0.05). Alertys Ruminant Pregnancy test was a reliable for pregnancy detection in sheep on Day 35 after artificial insemination.

Relationships between follicle and corpus luteum size and vascularization with ovulation, progesterone production, and pregnancy in Nellore beef cattle

ABSTRACT The objective of this study was to evaluate the relationships between preovulatory follicle (POF) and corpus luteum (CL) diameters, and POF and CL vascular perfusion with progesterone production, ovulation, and pregnancy in Nellore cows subjected to timed artificial insemination (TAI). Nellore cows (n = 201) were subjected to ovulation synchronization and later to ultrasound evaluation of POF and CL at the time of insemination (D0) and seven days later (D7), respectively. Females were divided into three categories according to the POF diameter assessed at the time of insemination: small (SF), medium (MF), and large (LF) follicles. The LF group had a greater number and intensity of pixels in the POF ultrasound exam compared with the SF group. The CL flow intensity and progesterone concentration were also higher in the LF group. The SF group showed lower flow intensity and lower ovulation rate compared with the others. When non-pregnant females were compared to pregnant ones, no difference was observed in any of the analyzed variables. The results show for the first time in Nellore cattle the relationship between the size of ovarian structures and blood flow (quantity and intensity) as well as the ability of the CL to produce progesterone. The intensity of the POF pixels proved to be relevant, demonstrating correlations with the size and flow of the CL, which were not found when evaluating only the number of pixels, thus revealing the importance of evaluating complementary characteristics of the flow.

Association between chronic Anaplasma marginale and Babesia spp. infection and hematological parameters of taurine heifers / Associação entre infecção crônica por Anaplasma marginale e Babesia spp. e parâmetros hematológicos de novilhas taurinas

The aim of this study was to investigate the association between chronic Anaplasma marginale and Babesia spp. infection and hematological parameters of pregnant and non-pregnant taurine heifers. Blood samples from 94 females were collected on the first day (D-10) of timed artificial insemination (TAI) protocol and on pregnancy diagnosis (D+34). Hematological parameters were determined and compared between pregnant (PG) and non-pregnant (NPG) heifers, and within group at different sampling days. Real-time PCR (qPCR) was used to determine A. marginale and Babesia bovis infection, and for absolute quantification of Babesia spp. between PG and NPG groups. Correlation analysis was performed between the number of gDNA copies (CN) of Babesia spp. and hematological parameters. On D-10, mean hemoglobin concentration was higher for NPG, and hematocrit and total plasma protein were higher on D+34 for both groups. There was no difference in Babesia spp. CN between groups. In the first qPCR, all heifers were positive for A. marginale and B. bovis. Significant correlations were found between hemoglobin and erythrocyte and between hemoglobin and hematocrit (r = 0.8082 and r = 0.3009, respectively). Low levels of A. marginale and Babesia spp. did not affect hematological parameters of chronically infected pregnant and non-pregnant taurine heifers.(AU)

O objetivo deste estudo foi investigar a associação entre infecção crônica por Anaplasma marginale e Babesia spp. e parâmetros hematológicos de novilhas taurinas prenhes e não prenhes. Sangue de 94 fêmeas foi coletado no primeiro dia (D-10) do protocolo de inseminação artificial em tempo fixo (IATF) e no diagnóstico de gestação (D+34). Parâmetros hematológicos foram comparados entre novilhas prenhes (PG) e não prenhes (NPG) e dentro dos grupos entre dias de coleta. Usando-se PCR em tempo real (qPCR), determinou-se a infecção por A. marginale e Babesia bovis e quantificação absoluta de Babesia spp. Entre os grupos PG e NPG. A análise de correlação foi realizada entre o número de cópias (CN) de Babesia spp. e parâmetros hematológicos. No D-10, a concentração de hemoglobina foi maior para NPG e hematócrito, e proteína plasmática total foram maiores em D+34 para ambos os grupos. Não houve diferença para CN de Babesia spp. entre os grupos. Na primeira qPCR, todas as novilhas foram positivas para A. marginale e B. bovis. Correlações significativas foram encontradas entre hemoglobina/eritrócito e hemoglobina/hematócrito (r=0,8082 e r=0,3009, respectivamente). Baixos níveis de A. marginale e Babesia spp. não afetaram os parâmetros hematológicos de novilhas taurinas prenhes e não prenhes cronicamente infectadas.(AU)

Occurrence of Cryptosporidium in Anglonubian goats in the municipality of Teresina, state of Piauí, Brazil / Ocorrência de Cryptosporidium em cabras da raça Anglonubiana no município de Teresina, estado do Piauí, Brasil

This research aimed to investigate the occurrence of Cryptosporidium and correlate it with types of housing, feces consistency, and physiological parameters related to the reproductive status of Anglo-Nubian goats reared in the State of Piauí, Brazil. A total of 180 fecal samples were collected from 60 non-pregnant and lactating goats with a mean weight of 35 kg, a body condition score of 3.5, and a mean age of three years from an experimental herd at the Federal University of Piauí (UFPI). Oocysts of protozoa of the genus Cryptosporidium could be found in the studied animals using the modified Ziehl-Neelsen technique in fecal smears and the image analysis system to perform morphometry. Each independent variable in the quantitative and qualitative analyses, that is, weight, body condition score (BCS), physiological status (non-pregnant or lactating), feces consistency (normal, pasty, or diarrheal), and floor types (concrete and slatted), was tested with the dependent variable (positive samples, i.e., the presence of Cryptosporidium oocysts). Twenty-four out of the total number of fecal samples were considered positive for the presence of the protozoan, which means that 13.3% of the animals were parasitized. Moreover, 100% of the positive feces samples had normal consistency (firm) and all parasitized animals were reared in pens with a concrete floor. A statistical variation was observed in the BCS of parasitized animals compared to non-parasitized ones (p > 0.0253). The results showed that the occurrence of Cryptosporidium in experimental goats located in the municipality of Teresina, State of Piauí, Brazil, was considered low, requiring sanitary management measures to prevent infection in animals and humans. This is the first report of Cryptosporidium infection in goats in the State of Piauí.

O objetivo desta pesquisa foi investigar a ocorrência de Crypstoporidium e correlacionar com tipos de alojamento, consistência das fezes e parâmetros fisiológicos ligados ao estado reprodutivo de cabras da raça Anglonubiana criadas no estado do Piauí, Brasil. Foram utilizadas 180 amostras de fezes de 60 cabra, com peso médio de 35kg, escore de condição corporal de 3,5, com idade em média de três anos, e cabras vazias e lactantes, de um rebanho experimental da Universidade Federal do Piauí (UFPI). Utilizando-se a técnica de Ziehl-Neelsen modificada em esfregaço fecal e sistema de análise de imagens para a realização da morfometria, foi possível encontrar oocistos de protozoários do gênero Crypstoporidium nos animais estudados. Nas análises quantitativa e qualitativa, cada variável independente: peso, escore de condição corporal (ECC), estado fisiológico (vazia ou lactante), consistência das fezes (normal, pastosa ou diarreica) e tipos de piso (concreto e ripado), foi testada com a variável dependente (amostras positivas, ou seja, presença de oocistos de Cryptosporidium). Do total de amostras fecais analisadas, 24 delas foram consideradas positivas à presença do protozoário, o que significa que 13,3% dos animais estavam parasitados na ocasião da pesquisa. Foi observado que 100% das amostras de fezes positivas apresentaram consistência normal (firme) e que todos os animais parasitados eram criados em aprisco com piso de concreto. Houve uma variação estatística no ECC dos animais parasitados comparados aos não parasitados (p > 0,0253). Os resultados evidenciaram que a ocorrência de Cryptosporidium em caprinos experimentais localizados no município de Teresina, no estado do Piauí, foi considerada baixa, sendo necessária medidas de manejo sanitário para prevenir a infecção nos animais e no homem. Este é o primeiro relato da infecção por Cryptosporidium em cabras no estado do Piauí.

The in vitro effects of n-3 fatty acids on immune response regulation of bovine ex vivo endometrial explants / Os efeitos in vitro dos ácidos graxos n-3 na regulação da resposta imune de explantes endometriais bovinos ex vivo

Fatty acids are considered metabolic intermediaries, although new facts indicate they also work as signaling molecules with different roles in the immune response. Based on that, in this study, we investigated the anti-inflammatory effects of n-3 polyunsaturated fatty acids (PUFAs) as eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and α-linolenic acid (LNA) in ex vivo bovine endometrial explants. For this, two groups were formed: (1) LPS-challenged and (2) control, both to evaluate the accumulation of proinflammatory cytokines as interleukin 1ß (IL1B) and interleukin 6 (IL6). To develop the study, bovine female reproductive tracts from non-pregnant Angus heifers without evidence of reproductive diseases were selected. Endometrial explants were processed and treated for 24 h with EPA, DHA, and LNA in five different concentrations (0µM, 50µM, 100 µM, 200µM and 400 µM) and then, challenged with LPS for 24 h. Supernatants were collected to evaluate the concentration of IL1B and IL6 by ELISA. Explants treated with EPA from control groups reduced the concentrations of ILB (200µM) and IL6 (400 µM), and IL6 (50 µM; 100 µM) from the LPS-challenged group. DHA decreased the accumulation of IL1B and IL6 at 200 µM on explants from the LPS-challenged group, and 200 µM reduced IL6 from the control group. In contrast, explants treated with LNA only reduced the accumulation of IL1B to 400µM (from both groups). In conclusion, the EPA acid is the best anti-inflammatory option to decrease the concentration of both pro-inflammatory cytokines (IL1B and IL6) from LPS-challenged and control groups in bovine endometrial explants; while LNA evidence to be the last option to promote an anti-inflammatory response.(AU)

Os ácidos graxos são considerados intermediários metabólicos, embora novos fatos indiquem que eles também atuem como moléculas sinalizadoras com diferentes papéis na resposta imune. Dessa forma, este estudo investigou os efeitos anti-inflamatórios de ácidos graxos poliinsaturados n-3 (PUFAs) como ácido eicosapentaenóico (EPA), ácido docosahexaenóico (DHA) e ácido α-linolênico (LNA) em explantes endometriais ex vivo de bovinos. Para tal, o experimento foi divido em dois grupos: (1) Desafiado-LPS e (2) Controle, para que então pudesse avaliar o acúmulo de citocinas pró-inflamatórias como interleucina 1ß (IL1B) e interleucina 6 (IL6). Foram selecionados tratos reprodutivos de fêmeas bovinas de novilhas Angus não prenhes sem evidência de doenças reprodutivas. Explantes endometriais foram processados e tratados por 24h com EPA, DHA e LNA em cinco concentrações diferentes (0µM, 50µM, 100 µM, 200µM e 400 µM) e, em seguida, desafiados com LPS por mais 24h. Os sobrenadantes foram colhidos para avaliar a concentração de IL1B e IL6 pelo teste de ELISA. Os explantes tratados com EPA dos grupos de controle reduziram as concentrações de ILB (200 µM) e IL6 (400 µM) e no grupo desafiado com LPS houve redução das concentrações de IL6 (50 µM; 100 µM). Nos explantes do grupo desafiado com LPS, o DHA diminuiu o acúmulo de IL1B e IL6 nas concentrações de 200 µM, e no grupo controle reduziu IL6 nas concentrações de 200 µM, enquanto os explantes tratados com LNA reduziram apenas o acúmulo de IL1B a 400 µM (de ambos os grupos). Em conclusão, o ácido EPA provou ser a melhor opção anti-inflamatória para diminuir a concentração de ambas as citocinas pró-inflamatórias (IL1B e IL6) de grupos desafiados com LPS e controle em explantes endometriais bovinos; enquanto o LNA evidencia ser a opção menos viável para promover uma resposta anti-inflamatória.(AU)

Use of new recombinant proteins for ovarian stimulation in ruminants

Currently, gonadotropin products (follicle stimulating hormone, FSH, and luteinizing hormone, LH) used in animal reproduction are produced by extraction and purification from abattoir-derived pituitary glands. This method, relying on animal-derived materials, carries the potential risk of hormone contamination and pathogen transmission. Additionally, chorionic gonadotropins are extracted from the blood of pregnant mares (equine chorionic gonadotropin; eCG) or the urine of pregnant women (human chorionic gonadotropin; hCG). However, recent advancements have introduced recombinant gonadotropins for assisted animal reproduction therapies. The traditional use of FSH for superovulation has limitations, including labor requirements and variability in superovulation response, affecting the success of in vivo (SOV) and in vitro (OPU/IVEP) embryo production. FSH treatment for superstimulation before OPU can promote the growth of a homogenous follicular population and the recovery of competent oocytes suitable for IVEP procedures. At present, a single injection of a preparation of long-acting bovine recombinant FSH (rFSH) produced similar superovulation responses resulting in the production of good-quality in vivo and in vitro embryos. Furthermore, the treatment with eCG at FTAI protocol has demonstrated its efficacy in promoting follicular growth, ovulation, and P/AI, mainly in heifers and anestrous cows. Currently, treatment with recombinant glycoproteins with eCG-like activity (r-eCG) have shown promising results in increasing follicular growth, ovulation, and P/AI in cows submitted to P4/E2 -based protocols. Bovine somatotropin (bST) is a naturally occurring hormone found in cows. Recombinant bovine somatotropin (rbST), produced through genetic engineering techniques, has shown potential in enhancing reproductive outcomes in ruminants. Treatment with rbST has been found to improve P/IA, increase donor embryo production, and enhance P/ET in recipients. The use of recombinant hormones allows to produce non-animal-derived products, offering several advantages in assisted reproductive technologies for ruminants. This advancement opens up new possibilities for improving reproductive efficiency and success rates in the field of animal reproduction.(AU)

Maternal and fetal ultrasonography, vulvar temperature and vaginal mucous impedance for the prediction of parturition in Saanen does

The aim of this study was to evaluate and correlate modifications of vaginal mucous impedance, vulvar temperature and ultrasonographic measurements (echobiometric parameters) to parturition in pregnant Saanen does. 30 does were selected for the study and submitted to an estrus synchronization protocol and natural mating. The females were evaluated daily from Day 143 of pregnancy to parturition. For the sonographic evaluations, the following structures were measured: biparietal diameter, thoracic diameter, abdominal diameter, ocular orbit, kidney length, kidney height, cardiac area, placentome length, cervical measurement and fetal heart rate; by means of two different approaches: transrectal and transabdominal, using a 7.5 MHz linear transducer. The vaginal mucous impedance was assessed using an electric estrous detector and vulvar temperature was measured using a non-contact infrared thermometer. Statistical analysis was performed using the R-project software and the significance level was set at 5% for all tests. 25 Saanen does became pregnant, resulting in 80.33% pregnancy rate. Fetal heart rate was negatively correlated to the hours to parturition (p<0,001; r-Pearson= -0,451), as well as vaginal temperature (p= 0,001; r-Pearson= -0,275), while cervical thickness was positively correlated to hours to parturition (p<0,001; r-Pearson= 0,490). The echobiometric parameters (biparietal diameter, thoracic diameter, abdominal diameter, ocular orbit, kidney length and height, cardiac area, placentome length), as well as vaginal mucous impedance did not vary throughout the timepoints of evaluation and did not correlate to the moment of parturition. It was concluded that the parameters of fetal heartbeat, vaginal temperature and cervical effacement in the last week of pregnancy provide valuable information regarding the proximity of parturition.(AU)

Lesão facial grave em um gambá-de-orelha-branca (Didelphis albiventris) - tratamento com ozonioterapia / Severe facial wound in a white-eared opossum (Didelphis albiventris) - treatment with ozone therapy

Background: Ozone therapy (OT) is a type of treatment that uses ozone gas, which has high oxidizing power. OT has been utilized for various diseases and to accelerate the wound healing process and reduce inflammation. Additionally, OT is a low-cost treatment recommended for patients that can be easily restrained. There are several methods of ozone administration, including topical use through bagging, cupping, or rectal insufflation, in addition to ozonated autohemotherapy. Based on the benefits of OT and the scarcity of reports describing its effects on white-eared opossums (Didelphis albiventris), we report on the use of ozone in the treatment of a severe facial wound in a white-eared opossum. Case: An adult female white-eared opossum weighing 1.6 kg was rescued from a fire. The patient was pregnant, dehydrated, hypothermic and had an extensive burn wound in the facial region. Due to the stress of captivity and its aggressive behavior, the animal needed to be chemically restrained with dexmedetomidine and butorphanol. Following sedation, the cephalic vein was catheterized and intravenous fluid therapy with ringer lactate was started to treat dehydration. Next, enrofloxacin, meloxicam, and tramadol hydrochloride were administered. Body rewarming was performed with thermal bags. The microbial load of the wound was reduced with chlorhexidine degerming solution followed by washing with saline solution. A thin layer of silver sulfadiazine cream was applied to the wound area. Over the following days the patient was treated with OT using the cupping technique combined with ozonated sunflower oil. After the first applications there was an improvement in the appearance of the wound, which initially had a purulent aspect. After 20 days of treatment the wound showed good granulation and there were no signs of infection, and at 45 days of treatment it was completely healed. The patient was then released with her baby. Discussion: Ozone therapy was included in the patient's treatment plan to enhance the tissue repair process in order to reintroduce the animal to the wild in the shortest possible time. The strategy was successful in improving healing and reducing tissue inflammation and infection. Ozone is known to cause the inactivation of microorganisms as a result of the oxidation of phospholipids and lipoproteins present in the cell membrane of bacteria and to interrupt viral replication. Ozone helps in the perfusion of injured tissues, contributing to the entry of humoral components, which control inflammation. Moreover, ozone contributes to increase supply of oxygen and nutrients to tissues and to stimulate increased production of fibrinogen and formation of granulation tissue. Ozone increases the enzymatic antioxidant response, while also playing a role in immunoregulation. Ozone can be used in several ways and via various methods with potential routes including intravenous, urethral, intramuscular, subcutaneous, or intra-articular, in addition to rectal and intramammary insufflation. Immunomodulatory, anti-inflammatory, analgesic, and antiseptic effects can be achieved with different doses and concentrations, which enables its application in different types of pathologies. Because of the aggressive behavior of the animal, in the current case we opted for the cupping technique combined with ozonated oil, both techniques which can be administered easily with only physical restraint of the patient. Cupping allows for the provision of a higher concentration of ozone gas locally whereas the ozonated oil maintains the effect of the gas for slightly longer, enhancing its effect and promoting tissue repair. The present report showed that OT is an effective alternative for the treatment of burn wounds in opossums, without causing adverse effects to the patient or her baby.

Gestational follow-up in ewes - placentome size and fecal progesterone metabolite levels

Background: Transabdominal ultrasonography in sheep allows real-time diagnosis and monitoring of pregnancy as well as measurement of the size of placentomes, which are structures that connect the fetal and maternal portions, that increase as pregnancy progresses. Progesterone is involved in the maintenance of gestation, with high levels noted during pregnancy. In this context, it is hypothesized that measurements of placentome and progesterone could help in monitoring pregnancy in sheep. Therefore, the main objectives of this study included evaluating whether placentome size and fecal progesterone levels are associated with gestational age in ewes and whether measurement of the placentome and progesterone could be used as an estimate of gestational age. Materials, Methods & Results: A total of 63 pregnant adult (between 2 and 5 years of age) Santa Inês ewes were monitored in the city of Boa Vista, Roraima. All ewes were kept in the paddock with Panicum maximun cv. tanzania, mineral salt, and water ad libitum, supplemented with 200 g/animal/day of soybean residue. Ewes were subjected to ultrasonography every 21 days from day 42 of pregancy until term to monitor pregnancy. Ultrasonography was performed using a linear probe to measure the placentome, including length × width in centimeters. Also every 21 days, fecal samples were collected for measurement of progesterone level. Feces were collected directly from the rectal ampulla, stored individually in plastic bags, and frozen at -20ºC until processing. In the laboratory, these feces samples were freeze-dried and subjected to hormonal extraction with 80% methanol. Then, the levels of fecal progesterone metabolites were measured by enzyme immunoassay. Data was tabulated and submitted to statistical analyses, including descriptive stastistic, correlation, KruskalWallis test and Tukey's test. The levels of fecal progesterone metabolites increased significantly between the 3rd and 4th months of pregnancy, with a decrease close to delivery. Placentome size increased significantly between the 2nd and 3rd months of pregnancy. Placentome measurements did not correlate with progesterone levels and showed a weak correlation with gestational age. Therefore, we concluded that the measurements of placentome size and a single evaluation of fecal progesterone metabolites were not considered efficient methods for estimating gestational age. Discussion: Measurement of progesterone level in fecal samples is a non-invasive hormone monitoring method that provides values like that of blood levels and is less stressful for the animals at the time of sample collection. The wide physiological variation makes it difficult or even useless to use fecal progesterone, when is measured only once per animal, as an indicator of the presence of pregnancy. Despite this limitation, serial sampling is useful for monitoring of hormones during the pregnancy period and is an important tool for research purposes. Placentome size measurements correlated weakly with gestational age and did not correlate with progesterone level. Thus, the presence of placentomes is indicative of the presence of pregnancy but was considered inefficient for determining gestational age in sheep.

Evaluation of extruded roughage with different additives in sheep diet / Evaluation of extruded roughage with different additives in sheep diet

The purpose was to evaluate the effect of extruded roughage Foragge® with different additives on intake, digestibility nutrients and nitrogen balance of sheep. Twenty adults, non-pregnant ewes with average weight 68 kg were used. The treatments were extruded roughage with additives (essential oil, virginiamycin, unpurified inactive yeast, tannin and purified inactive yeast). The design was in randomized blocks. The means were contrasted by SNK test, and the fecal score was tested by the Kruskal Wallis test (1952), at 5% significance. There was no difference in the intake of dry matter, crude protein, nitrogen, water, water in relation to dry matter, neutral detergent fiber (NDF), and acid detergent fiber (p > 0.05). As well as the digestibility of dry matter, crude protein and neutral detergent fiber, the fecal weight, fecal nitrogen, retained nitrogen and nitrogen retained in relation to nitrogen ingested (p > 0.05). However, dry matter intake as a function of body weight and metabolic weight, urinary nitrogen, hemicellulose intake, and hemicellulose as a function of NDF, were higher in the Foragge Factor® treatment (p < 0.05). The inclusion of different additives in the extruded roughage improved nutritional parameters, without causing disturbances.  

The purpose was to evaluate the effect of extruded roughage Foragge® with different additives on intake, digestibility nutrients and nitrogen balance of sheep. Twenty adults, non-pregnant ewes with average weight 68 kg were used. The treatments were extruded roughage with additives (essential oil, virginiamycin, unpurified inactive yeast, tannin and purified inactive yeast). The design was in randomized blocks. The means were contrasted by SNK test, and the fecal score was tested by the Kruskal Wallis test (1952), at 5% significance. There was no difference in the intake of dry matter, crude protein, nitrogen, water, water in relation to dry matter, neutral detergent fiber (NDF), and acid detergent fiber (p > 0.05). As well as the digestibility of dry matter, crude protein and neutral detergent fiber, the fecal weight, fecal nitrogen, retained nitrogen and nitrogen retained in relation to nitrogen ingested (p > 0.05). However, dry matter intake as a function of body weight and metabolic weight, urinary nitrogen, hemicellulose intake, and hemicellulose as a function of NDF, were higher in the Foragge Factor® treatment (p < 0.05). The inclusion of different additives in the extruded roughage improved nutritional parameters, without causing disturbances.  

Effect of ovarian status on reproductive performance in dairy heifers inseminated with sexed semen

Background: The intensive reproductive management in the dairy farms requires inclusion of a large group of replacement heifers in the breeding program for a shorter period. In this aspect, a creation of effective estrus synchronization protocols with timed artificial insemination (TAI) by sexed semen and optimization of the current ones have a crucial role for obtaining high pregnancy rate. These protocols are beneficial, because they led to reduced interval to first AI, lack of need for estrus detection, and allow obtaining a large group of female calves for on time. Because of limited fertilizing potential of sexed spermatozoa this type of semen is applied mainly for heifers, as fertility is higher compared to lactating cows. The main objective of the present study was to investigate the effect of ovarian status on the reproductive performance in dairy heifers subjected to estrus synchronization and timed artificial insemination with sexed semen. Materials, Methods & Results: Forty-eight Holstein healthy heifers separated in 2 groups were subjected to PGF2α-GnRH pre-treatment and Ovsynch or PRID-5-day estrus synchronization protocol and timed artificial insemination (TAI) with sex-sorted semen, staring 6 days after end of the hormonal pre-treatment. The ovarian status (presence of follicles with or not a corpus luteum) of the heifers at the begin of the treatment, on day of TAI and pregnancy rate in different groups were determined and compared. Additionally, the ovarian status at the begin of the treatment, ovarian status and size of preovulatory follicle (PF) on day of TAI and total values for both groups according to reproductive performance (pregnant or non-pregnant) were also analyzed. On day of TAI the animals with PF and a lack of corpus luteum (CL) in both groups were more that those with PF and CL (39.3% and 30% vs. 60.7 % and 70%), with significant (P < 0.05) difference in PRID-5-day group. The pregnancy rate tended to be higher in PRID-5 day than Ovsynch treatment (65% vs. 35.7%). A higher percentage (100% and 67.9%) of the pregnant animals in both treatments had not CL on day of TAI, and the size of the PF (1.58 ± 0.12 cm and 1.64 ± 0.13 cm) was increased (P < 0.05). Similar effects of the ovarian status on reproductive performance were obtained after a comparison of the total values between pregnant and non-pregnant animals. The percentage of heifers with observed PF without CL on day of TAI was significantly greater (P < 0.05) in pregnant compared to non-pregnant group (91.3% vs. 40%). The opposite dependence was determined for the parameter presence of follicles and corpus luteum (8.7% vs. 60%; P < 0.05). Furthermore, the size of the PF measured immediately before TAI was increased in animals became pregnant (1.60 ± 0.12 cm vs. 1.34 ± 0.17 cm; P < 0.05). Discussion: The analysis of the obtained results showed that the ovarian status on day of TAI affects the reproductive performance in dairy heifers subjected to estrus synchronization and timed artificial insemination with sexed semen. Hormonal pre-treatment with onset of PRID-5-day protocol 6 days later and TAI with sex-sorted semen ensure acceptable pregnancy rate. The greater preovulatory follicle and a lack of corpus luteum before insemination provide significantly (P < 0.05) more pregnant animals, compared to the cases when CL is presented. Ultrasound determination of the ovarian function before insemination can be used in selection of heifers for TAI with sex-sorted semen.

Níveis de progesterona e os parâmetros reprodutivos no período periovulatório de vacas Nelore submetidas ao protocolo de IATF / Progesterone levels and reproductive parameters in the periovulatory period of Nellore cows under FTAI protocol

Background: Fixed Time Artificial Insemination (FTAI) has achieved a significant evolution in the last 18 years, however, despite the progress achieved by modern FTAI programs, the conception rates obtained are still low. Therefore, this study aimed to evaluate the interrelation between progesterone levels in the periovulatory period and reproductive parameters of Nellore cows submitted to an FTAI protocol. Materials, Methods & Results: On a random day, called day 0 (D0), 57 cows received a P4 device associated with the intramuscular (IM) application of 2.0 mg of estradiol benzoate. On D9, the P4 devices were removed and then were administered 500 µg of cloprostenol sodium IM; 0.6 mg of estradiol cypionate IM and 300 IUI of Equine Chorionic Gonadotrophin IM. Blood samples were collected for the determination of serum P4 concentrations on D9 and D11 of the protocol. The evaluations of follicular diameter (DFOL), follicular wall area (AFOL) and the vascularization area of the follicle wall (VFOL) were carried out on D11 using B-mode ultrasonography examination and colour Doppler, and then the artificial inseminations were performed. The evaluation of the corpus luteum diameter (CLD), of the total corpus luteum area (CLA), of the area of corpus luteum vascularization (CLV) and blood sampling for determination of postovulatory P4 levels (Post-P4) were performed on D24. For the analysis of the P4 concentration the chemiluminescence method was used, with a sensitivity of 0.1 ng/mL. According to the P4 concentrations on D11, cows were divided into 2 groups, LOW LEVELS OF P4 and HIGH LEVELS OF P4. The diagnosis of pregnancy was performed using transrectal ultrasonography on D45, at this point the cows were divided into 2 groups, PREGNANT and NON-PREGNANT. The correlation between DFOL and P4 dosage on D11 was moderate, negative and significant and between the AFOL and the serum P4 levels on D9, was moderate, negative and significant. As for the other correlations between follicular and luteal parameters and serum P4 levels, these were low to moderate, negative and not significant. Cows in the LOW LEVELS OF P4 group had significantly larger diameter and follicular areas than the cows in the HIGH LEVELS OF P4 group, the other follicular and luteal parameters showed no statistical difference. Of the total 57 cows that were inseminated, 30 cows became pregnant. Cows in the PREGNANT group had serum P4 levels on D9 equivalent to that obtained by the NON-PREGNANT group. However, at D11 the cows that became pregnant presented significantly lower serum P4 levels than cows that did not become pregnant. Discussion: The results of the interrelation between follicular parameters and P4 levels obtained in the present study, pointed out that the lower the levels of P4, the higher the follicular parameters, corroborating with other authors. Thus, larger preovulatory follicles provided high ovulation rates. Periovulatory serum P4 levels did not significantly affect the morphofunctional parameters of the CL. Such findings may be justified by high periovulatory P4 levels resulting from less efficient luteolysis, exert a negative effect on the results of FTAI protocols, because progesterone inhibits the release of LH pulses. It is concluded that lower periovulatory P4 levels established a favourable condition for follicular development and fertility, however, morphofunctional parameters of the corpus luteum were not affected.

Evaluation of extruded roughage with different additives in sheep diet

The purpose was to evaluate the effect of extruded roughage Foragge®with different additives onintake, digestibility nutrients and nitrogen balance of sheep. Twenty adults, non-pregnant ewes with average weight 68 kg were used. The treatments were extruded roughage with additives (essential oil, virginiamycin, unpurified inactive yeast, tannin and purified inactive yeast). The design was in randomized blocks. The means were contrasted by SNK test, and the fecal score was tested by the Kruskal Wallis test (1952), at 5% significance.There was no difference in the intake of dry matter, crude protein, nitrogen, water, water in relation to dry matter, neutral detergent fiber (NDF), and acid detergent fiber (p>0.05).As well as the digestibility of dry matter, crude protein and neutral detergent fiber, the fecal weight, fecal nitrogen, retained nitrogenand nitrogen retained in relation to nitrogen ingested (p>0.05).However, dry matter intake as a function of body weight and metabolic weight, urinary nitrogen, hemicellulose intake, and hemicellulose as a function of NDF, were higher in the Foragge Factor®treatment (p<0.05). The inclusion of different additives in the extruded roughage improved nutritional parameters, without causing disturbances.(AU)

Influence of management and lactation order in the serum concentrations of proteins, metabolites, minerals, and enzymes of Bonsmara breed cows / Influência da gestação e ordem de lactação nas concentrações séricas de proteínas, metabólitos, minerais e enzimas de vacas da raça Bonsmara

This study aimed to evaluate the influence of gestation and lactation order on serum concentrations of proteins, metabolites, minerals, and enzymes in first-, second-, and third-lactation order Bonsmara cows. The Bonsmara breed, originated in South Africa, stands out for its tolerance to heat and productive characteristics, such as meat quality, high fertility, sexual precocity, ease of calving, and good maternal ability. These characteristics make the breed a genetic alternative for crossbreeding with zebu breeds. Blood samples were collected from 34, 29, and 30 first-, second-, and third-lactation order cows, respectively, totaling 93 cows. The samples were processed in an automatic multichannel analyzer using Labtest Diagnóstica® kits. The Kruskal-Wallis test was chosen to compare the values between lactation orders. The Mann-Whitney test was used to compare pregnant and non-pregnant cows within lactation orders. Among the analyzed constituents, the lactation order significantly influenced only the serum albumin concentration (ALB) (p<0.0001). Gestation significantly influenced the A:G ratio (p=0.034) in third-lactation cows, and cholesterol (Chol) (p=0.004), triglycerides (TRI) (p<0.0001), inorganic phosphorus (iP) (p=0.033), iron (Fe) (p=0.001), aspartate aminotransferase (AST) (p=0.018), and alkaline phosphatase (ALP) (p=0.039) in second-lactation cows and the general group. Creatinine (Crea) (p<0.0001) was influenced only in the general group. Therefore, gestation and lactation order significantly influenced the concentration of several biochemical serum constituents in Bonsmara cows, especially second-lactation order cows.

Objetivou-se avaliar a influência da gestação e ordem de lactação nas concentrações séricas de proteínas, metabólitos, minerais e enzimas de vacas da raça Bonsmara de primeira, segunda e terceira ordem de lactação. A raça Bonsmara, originada na África do Sul, se destaca por sua tolerância ao calor, e caracteristicas produtivas, como a qualidade da carne, a alta fertilidade, precocidade sexual, facilidade ao parto e boa habilidade materna. Essas características tornam a raça uma alternativa genética para realização de cruzamentos com raças zebuínas. Foram colhidas amostras de sangue de 93 vacas, sendo 34 de primeira ordem de lactação, 29 de segunda ordem de lactação e 30 de terceira ordem de lactação. As amostras foram processadas em analisador automático multicanal, utilizando kits da Labtest Diagnóstica®. Para confrontar os valores entre as ordens de lactação optou-se pelo teste Kruskall-Wallis. Para comparar vacas gestantes e não gestantes dentro das ordens de lactação, foi utilizado o teste de Mann-Whitney. Dos constituintes analisados, a ordem de lactação influenciou significativamente apenas a concentração sérica de albumina (ALB) (p<0,0001). A gestação influenciou significativamente na relação A:G (p=0,034), nas vacas de terceira lactação, no colesterol (COL) (p=0,004), triglicérides (TRI) (p<0,0001), fósforo inorgânico (Pi) (p=0,033), ferro (Fe) (p=0,001), aspartato aminotransferase (AST) (p=0,018) e fosfatase alcalina (FAL) (p=0,039) nas de segunda ordem e no valor do grupo geral. A creatinina (Crea) (p<0,0001) foi influenciada somente no grupo geral. Conclui-se que a gestação e ordem de lactação influenciou significativamente na concentração de vários constituintes bioquímicos séricos de vacas da raça Bonsmara, em especial nas de segunda ordem de lactação.

Addition of calcitic seaweed in the diet of sows positively affects the number of live-born piglets and milk parameters

This study was conducted to assess the effects of maternal dietary calcitic seaweed (CSW) on performance and blood metabolites of sows, and on performance, blood metabolites, intestinal microbiota, and parameters of gastrointestinal tract and bone of litters. On d 21 (post-insemination), non-pregnant sows were removed from the trial, remaining 19 sows in control group (without CSW) and 16 sows receiving CSW. Then, a total of 35 sows were allocated in a randomized block design with two treatments: control diet with calcitic limestone plus dicalcium phosphate (CTL) or CTL plus 0.4% CSW. In gestation, sows were fed twice a day (07:00 and 15:00 h) to reach an intake of 2.5 kg animal−1 day−1 divided into two equal meals. On parturition day, sows were offered only 0.5 kg feed animal−1. Throughout lactation, sows were fed three times a day (≅7 kg animal−1 day−1). All diets were provided as mash. Results suggested that sows fed CTL had litters with lower body weight at birth compared with those fed CSW. Sows fed CSW had 14.28% more live-born piglets and lower stillborns. Piglets from sows fed CSW showed greater calcium concentration on d 14 after birth than those from sows fed CTL. Sows fed CSW showed better milk chemical composition and an increase of 27.16% in milk production compared with those fed CTL. Piglets from sows fed CSW had an increase in cecum content in the Enterobacteriaceae count. This study showed that adding 0.4% CSW in the diet of pregnant and lactating sows as an organic calcium source positively influences the number of live-born piglets and the percentage of stillborns. In addition, milk composition and production are also improved without affecting piglets' biological response.(AU)

Evaluation of extruded roughage with different additives in sheep diet

The purpose was to evaluate the effect of extruded roughage Foragge® with different additives on intake, digestibility nutrients and nitrogen balance of sheep. Twenty adults, non-pregnant ewes with average weight 68 kg were used. The treatments were extruded roughage with additives (essential oil, virginiamycin, unpurified inactive yeast, tannin and purified inactive yeast). The design was in randomized blocks. The means were contrasted by SNK test, and the fecal score was tested by the Kruskal Wallis test (1952), at 5% significance. There was no difference in the intake of dry matter, crude protein, nitrogen, water, water in relation to dry matter, neutral detergent fiber (NDF), and acid detergent fiber (p > 0.05). As well as the digestibility of dry matter, crude protein and neutral detergent fiber, the fecal weight, fecal nitrogen, retained nitrogen and nitrogen retained in relation to nitrogen ingested (p > 0.05). However, dry matter intake as a function of body weight and metabolic weight, urinary nitrogen, hemicellulose intake, and hemicellulose as a function of NDF, were higher in the Foragge Factor® treatment (p < 0.05). The inclusion of different additives in the extruded roughage improved nutritional parameters, without causing disturbances.(AU)

Clinical, sonographic and anatomopathological aspects of ovarian teratoma in a heifer / Aspectos clínicos, ultrassonográficos e anatomopatológicos do teratoma ovariano em uma novilha

Teratomas rarely occur in domestic species, especially in cattle. These tumors originate in fetal life and are characterized by rapid growth, which justifies their frequent detection in young animals. This study reported a case of ovarian teratoma in a heifer. On physical examination, the main signs identified were apathy, abdominal distention and tension, empty rumen, and mushy diarrhea. During rectal palpation, a mass was identified in the pelvic region, which was suggestive of cysts on ultrasound examination. The animal underwent laparotomy, followed by euthanasia due to a poor prognosis. At necropsy, a 54 x 43 x 52 cm (length x width x thickness) tumor was observed in the right ovary with multiple cystic areas, in addition to masses associated with multiple adhesions of the intestinal loops and peritonitis. On histopathology, muscle, cartilage, bone, nervous and epithelial tissue, glands, hair with follicles, were identified in the affected ovary. There was mixed inflammation and foci of necrosis observed with a complete absence of ovarian architecture in both the ovaries. Infiltrations were identified in the lymph nodes and mesenteric vessels. Glandular ducts were seen from the serosa to the intestinal mucosa. A locally infiltrative and expansile ovarian teratoma was diagnosed accordingly. It is considered that this kind of tumor can induce abdominal distension and absence of estrus in previously healthy, non-pregnant heifers.

Os teratomas são tumores raros nas espécies domésticas, sobretudo em bovinos. Esses tumores são caracterizados por crescimento rápido e origem durante a vida fetal, o que justifica seu diagnóstico frequente em animais jovens. O presente trabalho relata um caso de teratoma ovariano em uma novilha. No exame físico, os principais sinais identificados foram apatia, distensão e tensão abdominal, rúmen vazio e diarreia pastosa. Durante a palpação retal, uma massa foi identificada na região pélvica. Ao exame ultrassonográfico, a massa era sugestiva de cistos. O animal foi submetido a laparotomia, seguido de eutanásia devido ao mau prognóstico. Na necropsia foi observado um tumor de 54 x 43 x 52 cm (comprimento x largura x espessura) no ovário direito com múltiplas áreas císticas, além de massas associadas à múltiplas aderências das alças intestinais e peritonite. Na histopatologia foram identificados no ovário acometido a presença de tecidos muscular, cartilaginoso, ósseo, nervoso e epitelial, com glândulas, pelos e folículos. Havia inflamação mista e focos de necrose com ausência completa de arquitetura ovariana em ambos os ovários. Infiltrações foram identificadas em linfonodos e vasos mesentéricos. Ductos glandulares foram vistos desde a serosa até mucosa intestinal. Diagnosticou-se um teratoma ovariano localmente infiltrativo e expansivo. Considera-se que este tumor pode induzir distensão abdominal e ausência de estro em novilhas previamente sadias e não gestantes.

Hidropsia fetal em dois filhotes da raça Buldogue Inglês / Fetal hydrops in two English Bulldog puppies

A hidropsia fetal é uma anomalia congênita, caracterizada pelo acúmulo excessivo de fluidos no espaço extravascular do feto, sendo denominada anasarca, quando o excesso de fluidos ocorre de forma generalizada, causando edema no tecido subcutâneo. Dessa forma, o aumento exagerado no tamanho do feto é esperado, podendo resultar em uma distocia obstrutiva, durante o parto. Os cães da raça Buldogue são mais susceptíveis a essa enfermidade, devido às características braquicefálicas; portanto, deve-se redobrar os cuidados durante a gestação, como utilizar a ultrassonografia para avaliar periodicamente os fetos, com o objetivo de preservar a saúde, tanto da gestante, quanto dos fetos. Vale evidenciar que, geralmente, essa enfermidade possui prognóstico de reservado a grave. Este presente trabalho relata o parto de uma cadela de 3 anos de idade, da raça Buldogue Inglês, que deu entrada em uma clínica veterinária, na cidade de Ourinhos/SP, com aproximadamente 62 dias de gestação, fruto de inseminação artificial, estando em trabalho de parto há aproximadamente 5 horas. A partir da realização dos exames físico e ultrassonográfico, foi observado que dois dos seis fetos apresentavam edema generalizado, estavam sem reflexos e com batimentos cardíacos diminuídos e abafados. A paciente foi submetida a cesariana de emergência. Os filhotes hidrópicos vieram a óbito, após tentativa de tratamento. Diante do exposto, conclui-se que a utilização da ultrassonografia deve ser priorizada, não apenas para diagnóstico, mas para acompanhamento da evolução da gestação e dos fetos, permitindo a visibilização de possíveis fetos hidrópicos e o agendamento da cesariana, procedimento cirúrgico mais indicado na presença dessa anomalia fetal.

Fetal hydrops is a congenital anomaly characterized by excessive accumulation of fluids in the extravascular space of the fetus, being called anasarca when excess fluid occurs in a generalized way, causing edema in the subcutaneous tissue. Thus, an exaggerated increase in the size of the fetus is expected, which may result in obstructive dystocia during birth. Bulldog dogs are more susceptible to this disease due to their brachycephalic characteristics; therefore, care should be redoubled during pregnancy, such as using ultrasound to periodically evaluate the fetuses to preserve the health of both pregnant and fetuses. It is worth noting that this disease usually has a reserved to severe prognosis. This present work reports the birth of a 3-year-old female English Bulldog that was admitted at a veterinary clinic in the city of Ourinhos/SP, with approximately 62 days of gestation, as a result of artificial insemination, being in labor for approximately 5 hours. From the physical and ultrasound examinations, it was observed that two of the six fetuses had generalized edema, had no reflexes, and presented reduced and muffled heartbeats. The patient underwent an emergency cesarean section. The hydropic puppies s died after an attempted treatment. Thus, it is concluded that the use of ultrasound should be prioritized not only for diagnosis, but for monitoring pregnancy evolution and fetuses, allowing the visualization of possible hydropic fetuses and scheduling of the cesarean section, which is the most indicated surgical procedure in the presence of this fetal anomaly.

Amputation of the front medial digit of a Holstein cow - successful parturition and milk production

Background: Diseases of the bovine digit and hoof remain major problems in dairy farming and the beef cow industry. Severe claudication accompanying swelling and pain at the lesion is often observed in deep digital sepsis, septic arthritis, severe sole ulcers, and chronic interdigital phlegmon. In addition, digital amputation is often performed in cases of white line disease or severe trauma, such as bone fractures. There are few reports of amputation of the medial digit of the bovine front hoof. In this case, amputation of the medial digit of the front leg of a pregnant cow was performed; its hospitalization and return to productivity is reported. Case: The bovine was a 9-year-old Holstein cow in the 6th month of pregnancy, weighing 671 kg. Upon admission, the cow could not bear weight on its right front leg, and swelling and a purulent discharge were observed in the coronary area of the medial digit. X-ray examination results indicated a periosteal reaction centered on the 3rd phalanx of the medial digit of its right front leg hoof and slight periosteal reactions at the adaxial sides of the middle and proximal phalanges, strongly suggestive of septic arthritis caused by infection. According to the X-ray examination results, no abnormalities of the lateral digit of the front right hoof were observed; thus, it was estimated that the post-amputation hoof would be able to bear weight. Therefore, a decision was made to perform immediate amputation of the medial digit of the right front leg hoof to reduce pain for improved delivery of the offspring and improved milk production during the lactation period, rather than allowing the lesion progress until the dry period or the expected date of delivery. According to the X-ray examination results, amputation of the 1st phalanx alone was expected to suffice for removal of the cause of the pain; thus, a decision was made to perform amputation at the edge adjacent to the 2nd phalanx. The right front medial digit was prepared for aseptic surgery, and infiltration anesthesia was performed. An incision was made with a surgical knife at the interdigit of the right front limb. A wire saw was inserted into the site to cut the 2nd phalanx in the anti-axial direction down to the skin to resect the lesion. White viscous pus was discharged at the resection site; therefore, the wound was lavaged with tap water and packed with povidone iodine-impregnated gauze. The wound was also covered with a diaper and dressed with non-elastic and elastic bandages. The dressing was changed daily to aid wound healing. No problems were observed in the standing-up motion or other relevant movements immediately after the surgery. Granulation tissue formed rapidly, approximately 2 weeks after the surgery, and the lesion dried gradually. Approximately one month after the surgery, the subject exhibited little difficulty in both standing up and walking. The subject successfully delivered its 7th offspring at the farm on postoperative day 93. Discussion: The animal of this study was a 9-year-old, pregnant cow, and although amputation of the front medial digit is a relatively rare procedure, the cow was able to deliver and return to production, owing to sufficient postoperative treatment and care. The case also demonstrated the advantage of X-ray examination in bovine hoof diseases for accurate diagnosis, precise operation, and prognostic assessment.

Frontiers in sheep reproduction - making use of natural responses to environmental challenges to manage productivity

This review addresses advances, directions and opportunities for research on sheep reproduction in the context of the global challenges of food security and climate change, and demand for 'clean, green and ethical' (CGE) animal management. The foundation of CGE management is an understanding of the physiological processes through which the reproductive system responds to changes in the animal's environment. These days, to the main environmental factors (photoperiod, nutrition, pheromones), we need to add stress from extreme weather events. With respect to nutrition in rams, we now have a deeper understanding of the responses of the brain centres that control gonadotrophin secretion (the kisspeptin system). At testis level, we have found that nutrition affects non-coding RNAs in Sertoli cells and germ cells, thus affecting the balance between cell proliferation and apoptosis. This proliferation-apoptosis balance is also affected during prenatal development, when undernutrition or stress in pregnant ewes seems to elicit epigenetic changes in developing gonads that could affect offspring fertility in adult life. With respect to nutrition in ewes, metabolic signals act directly on ovarian follicles, and thus change ovulation rate, but the variety of signals now includes the adipokines. An early concern was that nutritional supplements that increase ovulation rate would also increase embryo mortality but we now know that embryo survival is improved under field conditions. Finally, we had always thought that the efficiency gains from early puberty in lambs could only be achieved by accelerating fat accumulation, but we now know that faster muscle growth will achieve the same goal, offering two advantages in meat production systems. With respect to pheromones ('ram effect'), we have a deeper understanding of the brain responses (kisspeptin system) but, most importantly, a realization that the response of ewes to the ram signal involves cell division in memory centres. Many opportunities remain.(AU)