Alternative Traits for Genetic Evaluation of Mastitis Based on Lifetime Merit.

Genetic selection has achieved little progress in reducing mastitis incidence. Mastitis traits are problematic due to the lack of sensitivity of the data and reliance on clinical diagnosis, often missing subclinical cases, and/or on monthly somatic cell count (SCC) measurements. The current measure for mastitis is the lactation average of the somatic cells score (LSCS). We studied two datasets: (1) 148 heifers divided into non-intramammary infected, sub-clinically infected and clinical mastitis groups; (2) data from 89,601 heifers from Israeli Holsteins through the same period divided into "udder healthy" (UH) and "non-healthy" (UNH) by a threshold of SCC 120,000 cells/mL in all nine monthly milk recordings. In study 1, non-infected heifers had significantly (p < 0.05) more partum, production days and overall lifetime milk production compared to clinical and sub-clinically infected. In study 2, UH heifers (20.3%) had significantly higher (p < 0.01) lifetime milk, production days, and lactations. Subdividing datasets by sires, the same analyses detected differences in percentages of UH daughters between the sire groups. Lifetime milk production correlated (r = +0.83, p < 0.001) with udder health status. SCC threshold of less than 120,000 cells/mL during all first lactation measurements indicated healthy udder, providing a valuable insight that this dichotomous trait is advantageous for calculating lifetime net-merit index (NM$) over LSCS.

Economic Aspects of Bovine Ephemeral Fever (BEF) Outbreaks in Dairy Cattle Herds.

Bovine ephemeral fever virus (BEFV) is an arthropod-borne virus (arbovirus) transmitted by blood-feeding insects (mosquitoes and Culicoides biting midges). While the dispersal of arboviral diseases such as bovine ephemeral fever (BEF) into naive areas is often the result of globalization and animal movement, the endemization and local outbreaks of these diseases are mainly influenced by environmental changes. Climate change affects the activity, distribution, dynamics, and life cycles of these vectors (arthropods), the replication of viruses within their vectors, and weakens animal's immune systems. Although BEF does not currently occur in the Americas and Europe (other than in the western regions of Turkey), the risk of BEFV emergence, spread, and endemization in Europe is real. Over the past two decades, arboviruses such as the bluetongue virus (BTV) and Schmallenberg virus (SBV) have emerged in Europe without warning and caused significant losses to the dairy and meat industries. Since the European cattle population has never been exposed to BEFV, the economic losses to dairy and beef production in this continent due to the reduction in milk production, loss of valuable cows, and abortion, should BEF emerge, would probably be considerable. Moreover, arboviruses can also cause substantial financial damage due to restrictions on animal trade and transportation, like the current EHDV-8 outbreak in the Mediterranean basin. In this study, we used national data stored in the Israeli herd book to examine the economic aspects of BEF outbreaks in affected dairy cattle farms countrywide. Our results demonstrate that BEF outbreaks can have immediate and delayed effects, causing severe economic losses due to culling (loss of valuable cows) and a reduction in milk production that affects dairy farm income for months after clinical diagnosis. To our knowledge, this is the first extensive study on the impact of a BEF outbreak at a population level, enabling to conduct accurate risk assessments in future cases of BEFV emergence and re-emergence.

Evaluation of acoustic pulse technology as a non-antibiotic therapy for bovine intramammary infections: Assessing bacterial cure vs. recovery from inflammation.

Introduction: The spread of antimicrobial resistance (AMR) is a major threat to human and animal health. Therefore, new solutions are needed to prevent returning to a world without effective antibiotics. Mastitis in dairy cows is a major reason for antimicrobial use in food animal production, and mastitis-causing bacteria have the potential to develop AMR. In this study, acoustic pulse technology (APT) was explored as an alternative to antimicrobials for the treatment of mastitis in dairy cows. APT involves the local transmission of mechanical energy through soundwaves which stimulate anti-inflammatory and angiogenic responses in the udder. These responses promote udder recovery and enhance resistance to bacterial infections. Methods: We examined 129 Israeli dairy cows with mastitis in this prospective, controlled study to assess the efficiency of APT treatment on cure and recovery rates. An accurate diagnosis of suspected or confirmed infectious mastitis was made from cows having clinical signs of mastitis and/or somatic cell count (SCC) of above 400,000 cells/mL. The cows were divided into three groups: Group 1 (n = 29), cows with no bacterial findings (NBF); Group 2 (n = 82), cows with clinical signs of mastitis or SCC >400,000 cells/mL in the most recent test; and Group 3 (n = 18), cows with chronic mastitis (two or more tests with SCC >400,000 cells/mL within 3 months). All the cows received APT treatment, which involved 400 pulses on two sides of the infected quarter, delivered in three phases over 3 days. The cure for the mammary gland was indicated by the absence of bacterial growth in post-treatment cultures and recovery by a decrease in SCC to < 250,000 cells/mL in two of three post-treatment tests. Results and discussion: In Group 2, cure and recovery rates were 67.1 and 64.6%, respectively, and were not significantly different between Gram-negative and Gram-positive infections. A similar recovery rate was found in NBF cows. However, in cows with chronic mastitis, both the cure and recovery rates were significantly lower (22.2 and 27.8%, respectively). These results have important implications for dairy farmers, as APT treatment could lead to substantial savings of up to $15,106/year in a 100-cow herd, considering the national estimated prevalence of mastitis and the cost of individual treatment. APT should be further investigated as a viable and sustainable alternative to antimicrobial therapy for mastitis, offering economic benefits to dairy producers and the possibility of preventing AMR.

Estimating the Effect of the Kappa Casein Genotype on Milk Coagulation Properties in Israeli Holstein Cows.

In Israel, about 26% of produced milk is used to produce hard cheeses and 29% for soft cheeses. Milk with preferred coagulation properties requires a shorter coagulation time and yields a higher curd firmness than milk with inferior coagulation properties. Studies have shown that milk from cows with the B allele of kappa casein (κ-CN) produces more cheese than milk from those with A and E alleles. There is evidence that milk from AE or EE genotype cows is unsuitable for cheese production. In the early 1990s, the proportion of the B allele in Israeli Holstein cattle was about 17%, similar to its prevalence in the Holstein population worldwide. In recent years, however, its proportion has increased to about 40%. We analyzed milk coagulation properties as a function of the cow's κ-CN genotype, including time in minutes until the beginning of coagulation and curd firmness after 60 min-measured in volts via an optigraph device and scored on a scale of 0-4 by a laboratory technician. Cow selection was based on their sire's genotype, so that there would be sufficient genotypes that include the rare E allele. A total of 359 cows were sampled from 15 farms: 64 with genotype AA, 142 with AB, 41 with AE, 65 with BB, and 47 with BE. Data were analyzed via the general linear model procedure of SAS. We found the following: (a) There were significant differences between genotypes for optigraph-measured curd firmness. In a multi-comparison test, the BB genotype gave the highest curd firmness, and AB and BE showed a significant advantage compared to AA and AE (9.4, 8.6, 8.4, 6.9, 6.8 V, respectively). Assuming a frequency of about 55% for the A allele, about 30% of the milk delivered to dairy plants comes from AA cows. (b) There was a significant difference between the genotypes in technician-observed curd firmness, with BB scoring significantly higher than AA and AE. (c) The optigraph-measured curd firmness was significantly higher for milk from primiparous cows as compared to milk from second, third, or fourth lactation cows (8.9, 7.8, 7.9, 7.7 V, respectively). The technician-observed curd firmness was significantly higher for primiparous vs. multiparous cows. There was a clear advantage in curd firmness for genotypes that included the B allele compared to those with AA and AE genotypes. We can increase the proportion of the B allele in the population by insemination of cows using bulls with the genotypes AB and BB. This factor should therefore be included in the selection index.

Progesterone supplementation to improve fertility of selected subgroups of lactating cows during the summer and fall.

One major cause of low fertility of cows in the summer is progesterone deficiency. We found that insertion of a controlled intravaginal drug-releasing (CIDR) device containing progesterone after artificial insemination (AI) increases pregnancy per AI (P/AI) in cows with uterine disease and low body condition score after calving. Here, we treated only these two subgroups, during the summer and autumn. Control (n = 191 AI) and treatment (n = 230 AI) cows were inseminated at estrus and the treated group received a CIDR device on day 5 post-AI, for 14 days. Overall analysis of data during the summer and autumn indicated no significant differences between treatment and control groups. Analysis of the summer data only indicated a significant effect of treatment: P/AI was higher in CIDR-treated vs. control groups (34.2% vs. 19.3%; p < .038). Results indicated a 15% increase in P/AI during the summer for CIDR-treated cows in subgroups that had responded positively to the progesterone treatment.

Administration of GnRH at Onset of Estrus, Determined by Automatic Activity Monitoring, to Improve Dairy Cow Fertility during the Summer and Autumn.

We examined gonadotropin-releasing hormone (GnRH) administration at onset of estrus (OE), determined by automatic activity monitoring (AAM), to improve fertility of dairy cows during the summer and autumn. The study was performed on two dairy farms in Israel. The OE was determined by AAM recorded every 2 h, and a single im dose of GnRH analogue was administered shortly after OE. Pregnancy was determined by transrectal palpation, 40 to 45 d after artificial insemination (AI). Conception risk was analyzed by the GLIMMIX procedure of SAS. Brief visual observation of behavioral estrus indicated that about three-quarters of the events (n = 40) of visually detected OE occurred within 6 h of AAM-detected OE. Accordingly, the GnRH analogue was administered within 5 h of AAM-detected OE, to overlap with the expected endogenous preovulatory LH surge. Overall, pregnancy per AI (P/AI) was monitored over the entire experimental period (summer and autumn) in 233 first, second or third AI (116 and 117 AI for treated and control groups, respectively). Least square means of P/AI for treated (45.8%) and control (39.4%) groups did not differ, but group-by-season interaction tended to differ (p = 0.07), indicating no effect of treatment in the summer and a marked effect of GnRH treatment (n = 58 AI) compared to controls (n = 59 AI) on P/AI in the autumn (56.6% vs. 28.5%, p < 0.03). During the autumn, GnRH-treated mature cows (second or more lactations), and postpartum cows exhibiting metabolic and uterine diseases, tended to have much larger P/AI than their control counterparts (p = 0.07-0.08). No effect of treatment was recorded in the autumn in first parity cows or in uninfected, healthy cows. In conclusion, administration of GnRH within 5 h of AAM-determined OE improved conception risk in cows during the autumn, particularly in those exhibiting uterine or metabolic diseases postpartum and in mature cows. Incorporation of the proposed GnRH treatment shortly after AAM-detected OE into a synchronization program is suggested, to improve fertility of positively responding subpopulations of cows.

The Effect of Vaccination with Neospora caninum Live-Frozen Tachyzoites on Abortion Rates of Naturally Infected Pregnant Cows.

Neosporosis is a major cause of abortions in cattle worldwide. Recently a live attenuated vaccine showing promising results in preventing abortions, when administered at mid-pregnancy to seropositive cows, was developed. In this study, vaccination with 2 × 108 live frozen N. caninum tachyzoites (NcIs491) was used to immunize naturally infected seropositive pregnant dairy dams. The study was performed under field conditions in four herds, and a follow-up of three subsequent pregnancies was analyzed. A total of 1136 cows were serologically examined. Total seroprevalence was 41.4%, with 25.1% of the cows having titers of 1:800 or higher. Abortion rates were significantly higher in cows with high antibody titers (≥1:800) for two consecutive pregnancies. Vaccination was administered to 114 out of 285 cows with antibody titers higher than 1:800. Immunization resulted in lower abortion rates at three of the farms. Vaccine efficacy ranged from -19.8% to 75% at different farms, with overall efficacy of 28.4% in all four farms and overall efficacy of 58.2% in the three farms with positive results. Our results showed different vaccine efficacy in studied farms, suggesting that frozen live vaccination may generally be an effective method to control neosporosis in cattle.

Welfare Issues on Israeli Dairy Farms: Attitudes and Awareness of Farm Workers and Veterinary Practitioners.

Attitudes toward practical dairy cow welfare issues were evaluated based on a questionnaire answered by 500 dairy farm workers and 27 veterinary practitioners. Primarily, the effect of demographic characteristics on attitudes toward cattle welfare was tested. Professionally, five themes were identified: effect of welfare awareness on productivity, knowledge of cattle's senses and social structure, effects of man-animal interactions on milk yield, pain perception and prevention, and knowledge transfer from veterinary practitioners to farm workers. Farms with a higher welfare awareness score also had higher annual milk yield, with an annual mean difference of 1000 L of milk per cow between farms with higher and lower awareness scores. Veterinary practitioners showed high awareness of cows' social structure, senses, and pain perception. Farm workers were aware of the influence of man-animal interactions during milking and stress effects on milk yield, and the possible effect of man's behavior on heifers and cows. Practitioners and farm workers had different views regarding pain perception, mostly involving mutilation procedures. All veterinary practitioners advocated the use of pain alleviation in painful procedures, but only some of them instructed the farm workers to administer it. The survey results emphasize the variation in welfare knowledge and practical applications across farms, and the interest of both the animals and their managers to improve applied knowledge of best practice.

Physiological response of mammary glands to Escherichia coli infection:A conflict between glucose need for milk production and immune response.

The mammary immune and physiological responses to distinct mammary-pathogenic E. coli (MPEC) strains were studied. One gland in each of ten cows were challenged intra-mammary and milk composition (lactose, fat, total protein, casein), biochemical (glucose, glucose-6-phosphate (Glu6P), oxalate, malate, lactate, pyruvate and citrate, malate and lactate dehydrogenases, lactate dehydrogenase (LDH), nitrite, lactic peroxidase, catalase, albumin, lactoferrin, immunoglobulin) and clotting parameters were followed for 35 days post-challenge. Challenge lead to clinical acute mastitis, with peak bacterial counts in milk at 16-24 h post-challenge. Biochemical and clotting parameters in milk reported were partially in accord with lipopolysaccharide-induced mastitis, but increased Glu6P and LDH activity and prolonged lactate dehydrogenase and Glu6P/Glu alterations were found. Some alterations measured in milk resolved within days after challenge, while others endured for above one month, regardless of bacterial clearance, and some reflected physiological responses to mastitis such as the balance between aerobic and anaerobic metabolism (citrate to lactate ratios). The results suggest that E. coli mastitis can be divided into two stages: an acute, clinical phase, as an immediate response to bacterial infection in the mammary gland, and a chronic phase, independent of bacteria clearance, in response to tissue damage caused during the acute phase.

Oocyte maturation in plasma or follicular fluid obtained from lipopolysaccharide-treated cows disrupts its developmental competence.

Mastitis has deleterious effects on ovarian function and reproductive performance. We studied the association between plasma or follicular fluid (FF) obtained from endotoxin-induced mastitic cows, and oocyte developmental competence. Lactating Holstein cows were synchronized using the Ovsynch protocol. On Day 6 of the synchronized cycle, an additional PGF2α dose was administered, and either Escherichia coli endotoxin (LPS, 10 µg; n = 3 cows) or saline (n = 3 cows) was administered to one udder quarter per cow, 36 h later. Milk samples were collected and rectal temperatures recorded. Cows treated with LPS showed a typical transient increase in body temperature (40.3 °C ±â€¯0.4), whereas cows treated with saline maintained normal body temperature (38.9 °C ±â€¯0.04). A higher (P < 0.05) somatic cell count was recorded for cows treated with LPS. Plasma samples were collected and FF was aspirated from the preovulatory follicles by transvaginal ultrasound probe, 6 h after LPS administration. Radioimmunoassay was performed on plasma samples to determine estradiol and cortisol concentrations. Either FF or plasma was further used as maturation medium. In the first experiment, oocytes were matured in TCM-199 (Control) or in FF aspirated from cows treated with saline (FF-Saline) or LPS (FF-LPS). Cleavage rate to the 2- to 4-cell stage embryo did not differ among groups. However, the proportion of developed blastocysts on Day 7 postfertilization in the FF-LPS group tended to be lower for that in FF-Saline and was lower (P < 0.05) than that in the Control groups (10.6 vs. 22.4 and 24.4%, respectively). In the second experiment, oocytes were matured in TCM-199 (Control), or in plasma obtained from cows treated with saline (Plasma-Saline) or LPS (Plasma-LPS). Similar to the FF findings, cleavage rate did not differ among groups; however, the proportion of developing blastocysts tended to be lower in the Plasma-LPS group than in the Plasma-Saline group and was lower (P < 0.05) from that in the Control group (11.0 vs. 25.5 and 34.7%, respectively). The proportion of apoptotic cells per blastocyst, determined by TUNEL assay, did not differ among the experimental groups. The findings shed light on the mechanism by which mastitis induces a disruption in oocyte developmental competence. Further studies are required to clarify whether the negative effect on oocyte developmental competence is a result of LPS, by itself, or due to elevation of secondary inflammatory agents.

Seasonal and spatial variation in Culicoides community structure and their potential role in transmitting Simbu serogroup viruses in Israel.

Culicoides biting midges (Diptera: Ceratopogonidae) are a successful group of small (1-3 mm) haematophagous flies, some species of which are biological vectors of veterinary arboviruses, such as bluetongue virus, epizootic haemorrhagic disease virus, African horse sickness virus and Simbu serogroup viruses. In this study, we examine seasonal and spatial effects on the presence and distribution of Culicoides communities associated with ruminant and equine farms in Israel, and their infection with Simbu serogroup viruses. Our results demonstrate that both the vectors and the viruses are widely spread in Israel, including regions that were previously considered Culicoides-free. Moreover, our results show that although seasonality affects infection with Simbu serogroup viruses, both viruses and potential vectors can be found year round, suggesting continuous circulation of Simbu serogroup viruses in Israeli livestock farms. Finally, this study provides novel and basic information on Simbu serogroup-infected Culicoides in Israel: it demonstrates that Sathuperi, Shuni and Peaton viruses were circulating in Israel in 2015-2017 as they were found in C. imicola and C. oxystoma, both potential vectors of these viruses, and supplies the first-ever genomic detection of Sathuperi in Israel. Consequently, the data emerging from this study are of importance in understanding the epidemiology of arboviruses in Israel and are of relevance to the potential spread and possible future outbreaks of different Simbu serogroup viruses within the Mediterranean region and Europe.

Milk quality and milk transformation parameters from infected mammary glands depends on the infecting bacteria species.

The current study measured the influence of milk of subclinically infected glands by different bacteria species on the cow's milk. The effects of bacterial infection or inflammation on gland milk yield were related to the bacteria species that caused the infection. The volume of milk of the inflamed gland from the cow's milk yield was significantly lower (P<0.001) for the glands previously infected by Escherichia coli (PIEc) and those infected with Streptococcus dysgalactiae. Coagulation properties, rennet clotting time (RCT) and curd firmness (CF) also depended on the bacteria causing the infection. RCT values of all the inflamed glands were significantly longer (P<0.001) and CF values were significantly lower than that of the healthy ones. Moreover, in the whole milk, CF was also significantly lower and not proportional to the volume of the milk from the inflamed gland of the cow's milk. Calculation of the predicted 40% dry matter curd weight (PCW) on the cow level, including the healthy and inflamed glands or the healthy glands alone, showed that for 10 of 13 PIEc cows, the presence of the affected gland's milk in the whole cow milk resulted in a lower PCW value. Likewise, 7 of 20 cows infected by S. dysgalactiae had negative delta values. Unlike the latter bacteria, PCW from milk of glands infected with CNS increased, although in a lower magnitude than in the healthy glands. No correlation was found between logSCC in the whole cow milk (healthy and inflamed glands) and PCW.

Application of pancreatic phospholipase A2 for treatment of bovine mastitis.

Recent findings have indicated that secreted phospholipases A2 (sPLA2s) have anti-inflammatory functions, including relief of symptoms in a mouse model of mastitis. This prompted us to investigate the therapeutic application of sPLA2, PLA2G1B, for bovine mastitis. Initial testing of PLA2G1B's effect on bovine mammary epithelial cell (bMEC) line PS revealed no changes in cell viability or cytokine-secretion pattern. However, when cells were first treated with lipopolysaccharide endotoxin (LPS) or live bacteria (Escherichia coli or Staphylococcus aureus), incubation with PLA2G1B significantly improved cell viability, suggesting involvement of sPLA2s in protecting membranes from lipid-peroxidation damage, rather than a bactericidal action. When PLA2G1B was applied simultaneously with LPS, a significant short-term reduction in interleukin-8 secretion was observed compared with bMECs treated only with LPS, supporting previous reports that PLA2G1B affects interleukin-8 signaling in similar cells. Following the favorable outcome of the in vitro experiments, we tested PLA2G1B in vivo by mammary infusion into infected glands. In one of a small sample (n = 4) of lactating cows chronically infected with Streptococcus dysgalactiae, a single PLA2G1B treatment completely cleared inflammation and bacteria, demonstrating its potential to cure subclinical mastitis. PLA2G1B treatment did not affect coagulase-negative staphylococci infection. These types of mastitis may involve formation of a resistant biofilm, and its elimination may relate to sPLA2s' characteristic ability to aggregate with cellular debris, facilitating their internalization by macrophages. In a bovine model of clinical mastitis based on introduction of E. coli via the streak canal, a single mammary infusion of PLA2G1B led to faster recovery to pre-infection milk-yield levels and decrease of somatic cell counts. In this case, all of sPLA2s' modes of resolving inflammation may apply, including competitive binding of the sPLA2s' receptor, the inactivation of which confers resistance to endotoxic shock. Hence, this study strongly supports further research into PLA2G1B as a cure for bovine mastitis.

Experimental model of toxin-induced subclinical mastitis and its effect on disruption of follicular function in cows.

This study establishes an experimental model for subclinical mastitis induced by Gram-positive (G+) exosecretions of Staphylococcus aureus origin or Gram-negative (G-) endotoxin of Escherichia coli origin to examine its effects on follicular growth and steroid concentrations in Holstein dairy cows. Cows were synchronized with the Ovsynch protocol followed by a series of follicular cycles that included GnRH and PGF2α doses administered every 8 days. Cows received small intramammary doses of either G+ (10 µg, n = 10) or G- (0.5 µg, n = 6) toxin, or saline (n = 6; uninfected control) every 48 hours for 20 days. Follicular fluids were aspirated from preovulatory follicles before (aspiration one: control), at the end of (aspiration two: immediate effect), and 16 days after the end of (aspiration three: carryover effect) toxin exposure. During the 3 weeks of subclinical mastitis induced by G+ or G-, no local inflammatory signs were detected in the mammary gland and no systemic symptoms were noted: body temperatures of the treated cows did not differ from controls; plasma cortisol and haptoglobin concentrations were not elevated and did not differ among groups. Somatic cell count was higher in the treated groups than in controls, and higher in the G- versus G+ group. For analysis of reproductive responses, cows were further classified as nonaffected or affected based on an more than 20% decline in follicular androstenedione concentration in aspiration two or three relative to the first, control aspiration. Most G- (5/6) and 40% of G+ (4/10) cows were defined as affected by induced mastitis. An immediate decrease in the number of medium-size follicles was recorded on Day 4 of the induced cycle, toward the end of the 20-day mastitis induction, in the affected G+ compared with uninfected control group (1.0 ± 0.5 vs. 3.0 ± 0.4 follicles; P < 0.05); the affected G- and nonaffected G+ subgroups exhibited a similar numerical decline in the number of follicles. A carryover (but not immediate) decrease to 51% and 62% in follicular estradiol concentrations in G- affected group and G+ affected group was detected relative to controls (P < 0.05). The nonaffected G+ subgroup did not differ from its control counterparts. Based on the current experimental model, subclinical IMI induced by G+ or G- toxin disrupts follicular functions, and it seems that the ovarian pool of early antral follicles is susceptible to subclinical mastitis.

Induction of heparanase in bovine granulosa cells by luteinizing hormone: possible role during the ovulatory process.

Follicular development, follicular rupture, and corpus luteum (CL) formation are accompanied by extensive tissue remodeling. We examined whether heparanase (HPSE), which cleaves heparan sulfate glycosaminoglycans, is induced during these processes. Prostaglandin F2alpha injection, which initiated luteolysis and the development of a preovulatory follicle, moderately increased HPSE mRNA in bovine granulosa cells (GCs). GnRH, used to induce gonadotropin surge, markedly augmented HPSE mRNA levels 12 h after its injection. The temporal pattern of HPSE gene expression in follicular-luteal transition was further examined in follicles collected before, and 4, 10, 20, 25, and 60 h after GnRH injection. HPSE mRNA increased transiently 10-20 h after GnRH injection to levels 10-fold higher than in untreated heifers. HPSE protein levels were similarly elevated 20 h after GnRH injection in GCs, but not in the theca layer. Cyclooxygenase-2 (PTGS2) mRNA peaked before ovulation when HPSE levels returned to baseline levels. HPSE mRNA abundance also remained low in the CLs. The antiprogesterone, RU-486, elevated HPSE levels in GC culture, suggesting that progesterone secreted by CLs may inhibit HPSE. HPSE immunostaining was more abundant in GCs than thecae. In cultured GCs, LH induced a transient increase in HPSE mRNA 3-6 h after its addition, but not at 24 h. However, PTGS2 mRNA was clearly induced at this time. These findings suggest that: 1) HPSE may play a role in ovulation but much less so during CL development, and 2) GC-derived HSPE may be a novel member of the LH-induced extracellular matrix-degrading enzyme family and may contribute to follicular rupture.