Alfredo Quites Antoniazzi

Lack of FSH support enhances LIF–STAT3 signaling in granulosa cells of atretic follicles in cattle

Subordinate follicles (SFs) of bovine follicular waves undergo atresia due to declining FSH concentrations; however, the signalling mechanisms have not been fully deciphered. We used an FSH-induced co-dominance model to determine the effect of FSH on signalling pathways in granulosa cells of the second-largest follicles (SF in control cows and co-dominant follicle (co-DF2) in FSH-treated cows). The SF was smaller than DF in control cows while diameters of co-DF1 and co-DF2 in FSH-treated cows were similar. The presence of cleaved CASP3 protein confirmed that granulosa cells of SFs, but not of DFs and co-DFs, were apoptotic. To determine the effect of FSH on molecular characteristics of the second-largest follicles, we generated relative variables for the second largest follicle in each cow. For this, variables of SF or co-DF2 were divided by the variables of the largest follicle DF or co-DF1 in each cow. There was higher transcript abundance of MAPK1/3 and AKT1/2/3 but lower abundance of phosphorylated MAPK3/1 in SF than co-DF2 granulosa cells. Abundance of mRNA and phosphorylated protein of STAT3 was higher in granulosa cells of control SF than FSH-treated co-DF2. SF granulosa cells had higher levels of LIFR and IL6ST transcripts, the two receptors involved in STAT3 activation. Further, lower transcript abundance of interleukin 6 receptor (IL6R), another receptor involved in STAT3 activation, indicated that STAT3 activation in SF granulosa cells could be mainly due to leukemia inhibitory factor (LIF) signalling. These results indicate that atresia due to lack of FSH is associated with activated LIF-STAT3 signalling in SF granulosa cells, as FSH treatment reversed such activation.

Relationship between clinical and postmortem evaluation in repeat breeder beef cows

The objective of this study was to investigate the causes of the repeat breeder syndrome comparing clinical signs and postmortem findings in beef cows. The identification of factors affecting the reproductive tract can support decisions as to whether treatment of repeat breeder cows is justifiable than culling. Since all animals were submitted to clinical examination before being slaughtered, this study has a differential approach when compared with others, where genital tracts from abattoir were examined. In this study, 130 crossbred cows and heifers that have failed to conceive after three or more services were identified, submitted to a clinical examination and blood collection for karyotyping and sent to an abattoir. postmortem examinations included macroscopic evaluation of the genital tracts, bacteriology and histopathology of the uterus. Uterine alterations were predominant followed by oviduct and ovarian pathologies. Histopathological examination was more sensitive as a diagnostic tool than clinical examination. Repeat breeder cows had a predominance of uterine abnormalities (95%), such as inflammatory (42.9%) and degenerative (59.7%) conditions. Oviduct abnormalities were found in 29.8 % of animals. Furthermore, 1 out 10 karyotyped cows showed aneuploidy. Thus, this study stressed the importance of laboratory exams in the diagnosis of the causes of infertility of repeat breeder cows.

Intratesticular hypertonic sodium chloride solution treatment as a method of chemical castration in cattle

Castration of male calves is necessary for trading to facilitate handling and prevent reproduction. However, some methods of castration are traumatic and lead to economic losses because of infection and myiasis. The objective of the present study was to evaluate the efficiency of intratesticular injection (ITI) of hypertonic sodium chloride (NaCl; 20%) solution in male calf castration during the first weeks of life. Forty male calves were allocated to one of the following experimental groups: negative control-surgically castrated immediately after birth; positive control -intact males; G1-ITI from 1- to 5-day old; G2-ITI from 15- to 20-day old; and G3-ITI from 25- to 30-day old. Intratesticular injection induced coagulative necrosis of Leydig cells and seminiferous tubules leading to extensive fibrosis. Testosterone secretion and testicular development were severely impaired in 12-month-old animals from G1 and G2 groups (P<0.05), in which no testicular structure and sperm cells were observed during breeding soundness evaluation. Rectal and scrotal temperatures were not affected by different procedures. In conclusion, ITI of hypertonic NaCl solution induces sterility and completely suppresses testosterone secretion when performed during the first 20 days of life.

Função do interferon-tau durante o reconhecimento materno da gestação em ruminantes

Maternal recognition of pregnancy is the period when the conceptus signals its presence to the dam. In ruminants, it requires conceptus elongation, which coincides with maximum production of interferon-tau (IFNT). Conceptus IFNT acts in a paracrine manner silencing estrogen receptor alpha (ESR1) and oxytocin receptor (OXTR) in the luminal epithelium, thus preventing luteolytic prostaglandin F2 alpha (PGF2 ) pulses. Besides its role during maternal recognition of pregnancy, IFNT induces the expression of several interferon stimulated genes (ISGs) in the endometrium, corpus luteum (CL) and blood cells. Recently, it was suggested an endocrine role for IFNT during the period of maternal recognition of pregnancy in sheep. It was demonstrated that infusion of IFNT into the uterine vein can extend the estrous cycle beyond 32 days. This direct action of IFNT in extrauterine tissues induces ISGs expression, which might be involved in the rescue of the CL from the luteolytic effects of PGF2 pulses.

Corpus Luteum and Early Pregnancy in Ruminants

This review examines the function of the corpus luteum (CL) with emphasis on pregnancy in ruminant models and the possible impact of pregnancy in conferring luteal resistance to prostaglandin F2α (PGF2α). Critical processes involved with formation of the CL impact the capacity to secrete progesterone. Similarly, complete luteolysis is critically important in the event that pregnancy does not occur so that a new ovulation and opportunity for pregnancy is established. It is well known that serum progesterone must reach a critical nadir if ovulation and fertilization are to occur. Following fertilization, the function of the CL in providing adequate progesterone is critical in setting up an endometrial environment so that pregnancy is maintained. Benefits of supplemental progesterone during early pregnancy are inconsistent in ruminants. However, recent studies indicate that supplemental progesterone following artificial insemination (AI) may depend on the presence of the CL and the amount of progesterone released from the CL. The primary signal for maternal recognition of pregnancy, interferon-tau (IFNT), is secreted from the ruminant conceptus (embryo proper and extraembryonic membranes). IFNT disrupts release of PGF2α from the endometrium and is antiluteolytic through inhibiting uterine expression of the estradiol receptor (ESR1) or the oxytocin receptor (OXTR). Endocrine action of IFNT on peripheral blood mononuclear cells and on the CL may also contribute to immunomodulatory function and longer-term sustainability and function of the CL as pregnancy progresses.

Prostaglandin F2α-induced luteolysis involves activation of Signal transducer and activator of transcription 3 and inhibition of AKT signaling in cattle

Prostaglandin F2α (PGF) induces the precipitous loss of steroidogenic capabilities and cellular death in the corpus luteum of many species, yet the molecular mechanisms underlying this event are not completely understood. Signal transducer and activator of transcription 3 (STAT3) was activated in granulosa cells during follicle atresia, whereas AKT is immediately down‐regulated in the corpus luteum after PGF treatment in cattle; however, their involvement in both functional and morphological luteolysis in monovular species still need to be determined. Blood samples and corpus lutea were collected from cows before (0) and 2, 12, 24, and 48 hr after PGF treatment on Day 10 of the estrous cycle (4–5 cows per time point). Serum progesterone concentrations decreased by threefold (p < 0.05) within 2 hr, confirming functional luteolysis. The mRNA abundance of the pro‐apoptotic gene BAX increased 12–48 hr post‐PGF treatment (p < 0.05), while morphological luteolysis was observed 24 and 48 hr after PGF treatment, based on the loss of plasma membrane integrity, reduction of cytoplasmic volume, and pyknotic nuclei. Phosphorylated STAT3 increased, peaking at 12 hr, and remained elevated until 48 hr after PGF treatment. SOCS3 transcript abundance also increased (p < 0.05) starting at 2 hr post‐PGF treatment. In contrast, AKT phosphorylation decreased by 12 hr after treatment. Thus, activation of STAT3 and inactivation of AKT signaling are involved in structural regression of the corpus luteum.

Involvement of HPI-axis in anesthesia with Lippia alba essential oil citral and linalool chemotypes: gene expression in the secondary responses in silver catfish

In teleost fish, stress initiates a hormone cascade along the hypothalamus-pituitary-interrenal (HPI) axis to provoke several physiological reactions in order to maintain homeostasis. In aquaculture, a number of factors induce stress in fish, such as handling and transport, and in order to reduce the consequences of this, the use of anesthetics has been an interesting alternative. Essential oil (EO) of Lippia alba is considered to be a good anesthetic; however, its distinct chemotypes have different side effects. Therefore, the present study aimed to investigate, in detail, the expression of genes involved with the HPI axis and the effects of anesthesia with the EOs of two chemotypes of L. alba (citral EO-C and linalool EO-L) on this expression in silver catfish, Rhamdia quelen. Anesthesia with the EO-C is stressful for silver catfish because there was an upregulation of the genes directly related to stress: slc6a2, crh, hsd20b, hspa12a, and hsp90. In this study, it was also possible to observe the importance of the hsd11b2 gene in the response to stress by handling. The use of EO-C as anesthetics for fish is not recommended, but, the use of OE-L is indicated for silver catfish as it does not cause major changes in the HPI axis.

What is your diagnosis? Liver from a cow.

A 10-year-old Aberdeen Angus cow was presented to the Large Animal Medicine Service at Federal University of Santa Maria Veterinary Hospital with a history of weight loss for the past 6 months, watery diarrhea for the last 30 days, tenesmus, and fever. The owner reported the presence of the plant Senecio brasiliensis on pasture and that 10 cattle had died on his property during the past 4 years demonstrating similar antemortem clinical signs as the cow that was presented. On physical examination, the cow was dehydrated and mildly febrile (101.7°F) with pale mucous membranes. A moderate mature neutrophilia was noted on the CBC (17,300 cells/lL, RI 4000– 12,000). The biochemical serum panel revealed mild hypoalbuminemia (2.79 g/dL, RI 3.30–3.55), moderately decreased AST activity (36 IU/L, RI 78–132), markedly increased GGT activity (39.98 IU/L, RI 6.10–17.40), and moderately decreased urea concentration (35.33 mg/dL, RI 42.80–64.20). A fine-needle aspirate (FNA) smear of the liver was submitted for cytologic evaluation (Figure 1), and a liver biopsy was collected and routinely processed for histopathology. Figure 1. Fine-needle aspirate of liver from a cow with chronic signs of liver disease. Diff-Quik,940 objective.