Ovarian follicular cysts in dairy cows: an abnormality in folliculogenesis

Ovarian follicular cysts are a major reproductive problem in lactating dairy cows. The primary physiological defect leading to the formation of ovarian follicular cysts is a failure of the hypothalamus to trigger the preovulatory surge of luteinizing hormone (LH) in response to estradiol. The factor responsible for this hypothalamic defect may be progesterone. Intermediate levels of progesterone have been shown to prevent ovulation and promote persistence of dominant follicles in normal cycling cows. Recently, we found that 66% of cows with ovarian follicular cysts had progesterone concentrations in an unusual, intermediate range (0.1-1.0 ng/mL) at the time of their detection. A majority of new follicles (76%) that develop in the presence of these intermediate progesterone concentrations became cysts. Only 10% ovulated. Based on these observations, a novel model for the formation and turnover of ovarian follicular cysts is proposed.     

CONGENITAL FACIAL INFILTRATIVE LIPOMA IN A CALF

The clinical and pathologic findings related to an infiltrative facial lipoma in a 5-month-old female Holstein Friesian calf are reported. The tumor was congenital, increased in size with time, and deformed the left side of the face and cranial bones. The clinical condition of the calf was good; the only detectable abnormalities were poor weight gain and decreased motility of the left upper lip. Clinical and radiologic data indicated the mass was not removable. Muscle and lymph node invasion by tumor tissue were detected histologically. Infiltrative lipoma is a rare variety of lipoma that has been reported in dogs, cats, horses, and humans. Although its cytologic characteristics are those of a benign tumor, in this patient the invasiveness of the neoplasm was associated with poor prognosis.     

Effect of vaccination of cattle with the low virulence Nc-Spain 1H isolate of Neospora caninum against a heterologous challenge in early and mid-gestation

Live vaccines have emerged as one of the most potentially cost-effective measures for the control of bovine neosporosis. Previous studies have shown that Nc-Spain 1H is a naturally attenuated isolate of Neospora caninum and that immunisation with live Nc-Spain 1H tachyzoites generated a protective immune response in mice. The aim of this study was to evaluate the safety and efficacy of immunisation in cattle. N. caninum-seronegative heifers were immunised subcutaneously twice with 10⁷ live Nc-Spain 1H tachyzoites prior to artificial insemination. No adverse reactions or negative effects on reproductive parameters were recorded following immunisation. In immunised and non-challenged heifers, no foetal deaths were observed, and none of the calves was congenitally infected. The efficacy against N. caninum-associated foetal death and vertical transmission was determined after challenge with high doses of the Nc-1 isolate at 70 and 135 days of gestation, respectively. After the challenge in early gestation, the immunisation induced a protection of 50% against foetal death. In addition, the microsatellite analysis performed in PCR-positive tissue samples from foetuses that died after challenge infection showed that the profiles corresponded to the challenge isolate Nc-1. A degree of protection against vertical transmission was observed after challenge at mid-gestation; calves from immunised heifers showed significantly lower pre-colostral Neospora-specific antibody titres than calves from the non-immunised/challenge group (P < 0.05). Strong antibody and interferon gamma responses were induced in the immunised heifers. This study indicates that the immunisation before pregnancy with the Nc-Spain 1H vaccine isolate appeared to be safe and reduced the occurrence of N. caninum-associated abortion and vertical transmission in experimentally infected cattle. In light of these encouraging results, the next step for testing this live attenuated candidate should be the assessment of its efficacy and safety in naturally infected cattle.     

Cellular mechanisms by which oxytocin mediates uterine prostaglandin F2α synthesis in bovine endometrium: role of calcium

The objective of these experiments was to determine the role of Ca²⁺ during oxytocin-stimulated prostaglandin (PG) F_2α release from bovine endometrial tissue in vitro. Uteri were collected from dairy cows on the day after spontaneous luteal regression. Caruncular endometrial explants were dissected and incubated in vitro to determine phospholipase C activity or PGF_2α release. A23187 (a calcium ionophore) and maitotoxin (an activator of voltage-gated L-type calcium channels) stimulated release of PGF_2α in a concentration-dependent manner (P < 0.05). Thapsigargin (induces accumulation of Ca²⁺ in the cytoplasm by inhibiting endoplasmic reticulum Ca²⁺/ATPase pumps) stimulated release of PGF_2α in a concentration-dependent manner as well (P < 0.13). Oxytocin (10⁻⁶ M), AlF₄⁻ (a nonspecific activator of G-proteins; 10⁻⁵ M), A23187 (10⁻⁵ M), and melittin (a stimulator of phospholipase A₂; 10⁻⁴ M) stimulated PGF_2α release when explants were incubated in Ca²⁺-free medium (P < 0.10); however, oxytocin, A23187, or melittin were unable to stimulate PGF_2α release when explants were incubated in Ca²⁺-free medium containing the calcium chelator EGTA (P < 0.10). This treatment did not prevent oxytocin or AlF₄⁻ from stimulating phospholipase C activity (P < 0.08). CoCl₂ (a nonspecific Ca²⁺ channel blocker) and methoxyverapamil (a specific voltage-gated L-type Ca²⁺ channel blocker) prevented oxytocin from stimulating PGF_2α release (P < 0.05). Our results suggest that both extracellular and intracellular Ca²⁺ may be required for oxytocin to stimulate PGF_2α secretion in bovine endometrial tissue.     

Neospora caninum DNA detection by TaqMan real-time PCR assay in experimentally infected pregnant heifers

Neosporosis has been considered the main cause of abortion between the first and the second trimester of pregnancy in cattle. Therefore, the objective of this study was to identify the presence of Neospora caninum DNA obtained from experimental models based on the evaluation of different areas of the fetal nervous system and organs from heifers previously inoculated with NC-1 after or before insemination. This study was performed with Hereford × Nelore (n = 29) heifers and all animals were considered free of diseases at the beginning of the experiment. All animals were bred by fixed-time artificial insemination (TAI) and allocated as follows: (a) seronegative heifers subjected to TAI (TAI, n = 9), (b) heifers infected with N. caninun 60 days prior to TAI (NC-1 + TAI, n = 9), and (c) heifers submitted to TAI and infected with N. caninum 60 days later (TAI + NC-1, n = 11). The pregnancy was confirmed by transrectal ultrasonography 35 days after TAI and evaluated every 30 days until the end of gestation. Fetuses were collected surgically at 170 days of gestation, and immediately necropsied to remove tissues aseptically. Samples of the central nervous system (CNS), heart, kidney, lung, liver, skeletal muscle and caruncle were collected for DNA extraction. Days of gestation at abortion and interval from abortion to first insemination were examined by Student's t-test. At 35 days of gestation the pregnancy rates in the group NC-1 + TAI (4/9, 44.4%) was lower than in the control group (8/9, 88.8%, P < 0.05). At 60 days, the pregnancy rates in the NC-1 + TAI group (0/4, 0%) was lower compared to TAI + NC-1 (5/7, 71.4%) and control (6/8, 75.0%) groups (P < 0.05). Animals from the group NC-1 + TAI were re-inseminated 60 days after the first TAI. After pregnancy losses throughout the study, 5 animals (TAI), 3 animals (NC-1 + TAI) and 5 animals (TAI + NC-1) maintained pregnancy until 170 days of gestation. TaqMan RT-PCR demonstrated the presence of N. caninum DNA in the medulla and right posterior cortex in 3 out of 5 fetuses from the TAI + NC-1 group. We concluded that heifers infected after TAI had a higher incidence of the parasite at the fetus CNS. Identification of N. caninum by TaqMan RT-PCR would assist in the investigation of infection and in the evaluation of vaccines or therapeutic drugs to control neosporosis in cattle.     

Effect of neutral detergent soluble fibre on digestion, intestinal microbiota and performance in twenty five day old weaned rabbits

The effect of neutral detergent soluble fibre (NDSF) level on digestibility, fermentation traits, intestinal microbiota and performance was studied in weaned rabbits. A control diet (DA) containing 103 g NDSF/kg DM included dehydrated alfalfa as the main source of fibre. Another diet (B-AP) was formulated by replacing half of the dehydrated alfalfa with a mixture of beet and apple pulp resulting in 131 g NDSF/kg DM. A third diet (OH) was obtained by substituting half of the dehydrated alfalfa with a mix of oat hulls and a soybean protein concentrate and contained 79 g NDSF/kg DM. All diets contained similar levels of total fibre (NDSF + neutral detergent fibre), starch and protein (446, 208, and 199 g/kg DM, respectively). Forty-two rabbits (14/diet) weaned at 25 days were used to determine faecal digestibility from 32 to 35 days of age. This group, plus another nine rabbits/diet (23/diet), were used to determine weight of stomach and caecum and their contents, caecal fermentation traits and similarity rate (SR) of intestinal microbiota. Another 105 and 245 weaned rabbits were used to determine growth traits and mortality, respectively. From 25 to 39 days of age, rabbits were fed the experimental diets and from 39 to 60 days they were fed a commercial diet, including robenidine hydrochloride in all diets. Drinking water was supplemented with apramicine sulfate and tylosine tartrate throughout the experimental period. Faecal and energy digestibility increased linearly by 8% and NDF digestibility by 43% between extreme diets with NDSF inclusion (P < 0.001). Weight of total gastrointestinal tract decreased linearly and quadratically with NDSF reduction (P = 0.008 and P = 0.089, respectively). Stomach pH decreased linearly with increasing levels of NDSF (P ≤ 0.041). Weight of caecal contents increased linearly between animals fed OH and B-AP diets (P < 0.001). Level of inclusion of NDSF had no effect (P ≥ 0.12) on pH, VFA concentration and VFA molar proportions in caecal contents. Treatments appeared to influence the SR of caecal microbiota but a lesser effect was observed on ileal microbiota. Post weaning feed efficiency (25–39 days) increased linearly (P < 0.001) with NDSF inclusion by 10% between extreme diets, and by 3% in the whole fattening period (P = 0.027). Average feed intake during the post weaning (25–39 days) and the whole fattening period increased with NDSF reduction (P ≤ 0.079). No effect of NDSF was detected on average daily gain (P ≥ 0.15). Mortality decreased linearly with increasing levels of NDSF in the post weaning and in the whole fattening period (P = 0.086 and 0.016, respectively).     

Efficiency of nitrogen metabolism in beef cattle grazing pasture and supplemented with different protein levels in the rainy season

Supplementation of animals in the rainy season is essential to maximise weight gain. Thus, the objective was to evaluate the effects of supplementation with low-, medium- and high-protein levels on nutritional parameters of grazing beef cattle in the rainy season. Eight rumen-cannulated Nellore bulls (437 kg body weight) were used in a 4?×?4 double Latin square design with four supplements and four experimental periods. The treatments were mineral supplement (ad libitum) and supplements with low-protein (LPSU), medium-protein (MPS) and high-protein (HPS) levels, providing 106, 408 and 601 g/day of crude protein (CP), respectively. There was no difference (P?>?0.05) between treatments for dry matter and forage intake, ruminal pH, ruminal ammonia nitrogen (RAN) at time 0 (before supplementation) and microbial protein yield. Animals on MPS had a higher (P?>?0.05) intake and digestibility of CP, higher RAN levels (3 and 6 h after supplementation), increased nitrogen intake and a better nitrogen balance than animals on LPSU. The HPS provided higher (P?>?0.05) RAN at time 6 and a better nitrogen balance when compared with MPS. We therefore infer that the high-protein supplement positively impacts nitrogen metabolism and efficiency in grazing beef cattle during the rainy season.     

Host genotype and amoxicillin administration affect the incidence of diarrhoea and faecal microbiota of weaned piglets during a natural multiresistant ETEC infection

Enterotoxigenic Escherichia coli (ETEC) is the aetiological agent of postweaning diarrhoea (PWD) in piglets. The SNPs located on the Mucine 4 (MUC4) and Fucosyltransferase 1 (FUT1) genes have been associated with the susceptibility to ETEC F4 and ETEC F18, respectively. The interplay between the MUC4 and FUT1 genotypes to ETEC infection and the use of amoxicillin in modifying the intestinal microbiota during a natural infection by multiresistant ETEC strains have never been investigated. The aim of this study was to evaluate the effects of the MUC4 and FUT1 genotypes and the administration of amoxicillin through different routes on the presence of diarrhoea and the faecal microbiota composition in piglets naturally infected with ETEC. Seventy-one piglets were divided into three groups: two groups differing by amoxicillin administration routes—parenteral (P) or oral (O) and a control group without antibiotics (C). Faecal scores, body weight, presence of ETEC F4 and F18 were investigated 4 days after the arrival in the facility (T0), at the end of the amoxicillin administration (T1) and after the withdrawal period (T2). The faecal bacteria composition was assessed by sequencing the 16S rRNA gene. We described that MUC4 and FUT1 genotypes were associated with the presence of ETEC F4 and ETEC F18. The faecal microbiota was influenced by the MUC4 genotypes at T0. We found the oral administration to be associated with the presence of diarrhoea at T1 and T2. Furthermore, the exposure to amoxicillin resulted in significant alterations of the faecal microbiota. Overall, MUC4 and FUT1 were confirmed as genetic markers for the susceptibility to ETEC infections in pigs. Moreover, our data highlight that group amoxicillin treatment may produce adverse outcomes on pig health in course of multiresistant ETEC infection. Therefore, alternative control measures able to maintain a healthy faecal microbiota in weaners are recommended.     

The effect of different grazing heights on carcass and meat quality characteristics of Nellore cattle fattened in pastures of Convert grass

Carcass characteristic and meat quality from bulls and Nellore steers (n?=?64 total) subjected to different grazing heights (15, 25, 35, and 45 cm) were evaluated isolatedly, under continuous grazing and variable load, in pastures of Convert grass. The experimental period was from May 2015 to June 2017, in an area of 16 ha, divided into 16 paddocks of 1 ha. The experimental design was randomized blocks with four replications. Each paddock was grazed by three animals and regulators, used to adjust grazing heights. The final slaughter weight, hot carcass, and crude protein in the meat of steers were higher when the pastures were managed at 42 cm. When the pasture was managed between 25 and 35 cm in height, greater fat thickness, marbling, muscle:bone and muscle+fat:bone ratio and lower color* of the meat and percentage of bone were found. For the steers, the height of 40 cm provided higher fat thickness and marbling in the meat. The loss during thawing in meat was greater at 28 cm in height. The heights of grazing alter the carcass characteristics and meat of bulls and steers.

     

Relationship between serum acute phase protein concentrations and lesions in finishing pigs

To establish the relationship between serum levels of three acute phase proteins, haptoglobin (Hp), C-reactive protein (CRP) and serum amyloid A (SAA), and the occurrence and severity of lesions at slaughter, a study was carried out using 70 fattening pigs from a finishing unit. Pigs were divided into three groups: Group 1 (25 pigs with clinical signs of disease), Group 2 (25 apparently healthy pigs with lesions at slaughter) and Group 3 (20 apparently healthy pigs with no lesions at slaughter). Serum levels of CRP, SAA and Hp were significantly higher in pigs with clinical signs of disease than in apparently healthy animals. Additionally, in apparently healthy pigs, serum levels of Hp and CRP were significantly higher in animals with lesions than those without lesions. The extent and severity of lung lesions were related to serum levels of Hp.