Effects of mancozeb, metalaxyl and tebuconazole on steroid production by bovine luteal cells in vitro.

Mancozeb, metalaxyl and tebucanazole are widely used pesticides in agriculture and industry to treat plant pathogenic fungi. Livestock may be exposed to such substances by consuming contaminated plants. The present study was designed to evaluate the effects of these three fungicides on bovine luteal cell steroidogenesis. Luteal slices from mid-cycle corpus luteum were dissociated into single cell suspension in aerated (O2) culture media (DMEM/F12) by enzymatic digestion. The cells were incubated in newborn calf serum (10%) for 18 h and then with serum-free media containing mancozeb (0.01 µM, 0.1 µM, 1 µM), tebuconazole (1 µM, 10 µM, 100 µM) or metalaxyl (100 µM, 500 µM, 2500 µM) for additional 96 h. The medium was replaced on day 1 and 3; and the retrieved medium was stored at -20 °C until progesterone assay. Treatment of cells with three different fungicides induced dose dependent variable decrease in steroid synthesis during incubation periods. Incubation of cells with 1 µM mancozeb exhibited a 33% decline in steroid synthesis on day 3 and 48% decline on day 5 compared with controls. Treatment of cells with 100 µM tebuconazole and 500 µM metalaxyl resulted in a 65% and 31% decrease, respectively, in progesterone accumulation on day 5 of incubation. Fungicide induced suppressive effects on luteal steroidogenesis were as metalaxyl < tebuconazole < mancozeb. Results of the present study suggest that designated concentrations of all three fungicides studied might have varying degrees of adverse effects on luteal steroidogenesis.

Effect of ovarian activity on orthodontic tooth movement and gingival crevicular fluid levels of interleukin-1ß and prostaglandin E(2) in cats.

OBJECTIVE: To evaluate whether there is any correlation between ovarian activity and two potent bone-resorbing mediators (prostaglandin E(2) [PGE(2)], interleukin-1ß [IL-1ß]) secreted from the gingival crevicular fluid (GCF) during orthodontic tooth movement. MATERIALS AND METHODS: Eighteen female cats were included in this study. Animals were randomly divided into three groups (estrous, anestrous, and ovariectomized groups), each having six queens. Estrous was induced by administration of 150 IU equine chorionic gonadotropin (eCG) to queens of the estrous group. A closed-coil spring, applied with 80 g of tipping force to the canine, was attached between the maxillary canine and mini-implant. GCF was collected on days 0, 6, and 12 from each cat to examine PGE(2) and IL-1ß during orthodontic tooth movement in cats. The PGE(2) and IL-1ß levels were determined with enzyme-linked immunosorbent assay. RESULTS: There was no significant difference (P > .05) between anestrous and the ovariectomized groups in terms of tooth movement on days 6 and 12 of distalization. In contrast, tooth movement in the estrous group was lower (P < .05) than in the remaining two groups (anestrous and ovariectomized). The mean PGE(2) and IL-1ß levels of the canine teeth of the estrous groups were significantly lower than the remaining two groups on days 6 and 12 (P < .05) of coil spring applications. CONCLUSIONS: These results indicate that ovarian activity can affect orthodontic tooth movement and GCF levels of IL-1ß and PGE(2) in cats.

Effects of cholesterol and cAMP on progesterone production in cultured luteal cells isolated from pseudopregnant cat ovaries.

The present study was designed to incubate luteal cells isolated from pseudopregnant cats and to investigate the effects of cholesterol and cAMP on luteal progesterone production. Corpora lutea were collected from the cats on days 10 and 15 of pseudopregnancy. Luteal cells were isolated from the ovaries by collagenase digestion. Steroidogenic luteal cells were stained for 3beta-hydroxysteroid dehydrogenase (3beta-HSD) activity. Cells (2 x 10(4)) staining positive for 3beta-HSD were cultured for up to 7 days. The cells were treated with 22(R)-hydroxycholesterol (22R-HC) and dibutyryl cyclic AMP (dbcAMP) on days 1, 3 and 7. Treatment of cells with 22R-HC resulted in a dose-dependent increase (p<0.001) in progesterone production. When 22R-HC was used at a concentration of 10 microg/ml, it resulted in 2.7- and 5.1-fold increases in progesterone production on days 3 and 5, respectively. When the dose was doubled (20 microg/ml), treated cells produced four times more progesterone on days 3 and 7, and three times more on day 5. By day 7, progesterone production increased up to 9.1 times more than the control. Incubation of cells with both concentrations of dbcAMP (0.1 mM and 1 mM) resulted in significant stimulations of progesterone on days 5 and 7 (p<0.001). However, on day 3, only higher doses of dbcAMP (1 mM) resulted in significant stimulation (p<0.05). Progesterone production was increased up to 2- and 2.9-fold of the control when cells were treated with lower concentration of dbcAMP (0.1 mM) on days 5 and 7, respectively. Incubation of cells with 1 mM concentrations of dbcAMP induced a 3.2-fold increase on day 5 and a 5-fold increase on day 7. In conclusion, a successful incubation was performed for long-life culturing of luteal cells collected from pseudopregnant cats. The method works well and allows for optimal growth and development of cells in the culture. The present study also demonstrated that incubating cat luteal cells with 22R-HC and dbcAMP induces a significant increase in luteal progesterone synthesis.

Titres of alloantibodies against A and B blood types in non-pedigree domestic cats in Turkey: assessing the transfusion reaction risk.

The severity of a transfusion reaction depends on alloantibody titres within the recipients' blood. Determination of an agglutination titre of naturally occurring alloantibody may help to assess the risk of transfusion reactions following an unmatched transfusion in a cat population. In this group of 312 cats 227 had blood type A, 78 had blood type B, and seven had type AB blood. All type B cats tested showed gross evidence of agglutinating anti-A antibody with plasma titres ranging from 2 to 256. Among the 227 type A domestic cats tested for plasma anti-B alloantibody titres, 70% had gross agglutination with titres ranging from 2 to 16, while 17.6% had microscopic agglutination. The remaining 12.4% of the type A cats were negative for both gross and microscopic agglutination. Based on agglutinating titres, the relative risk of a transfusion reaction when type A or AB blood was given to a type B cat was 6.4% with acute severe reaction, acute mild reactions in 85.9% and premature red cell destruction in 7.7%. On the other hand, transfusion of type AB blood or type B blood to type A cats carries a potential risk of acute mild transfusion reaction in 4.4% and premature red cell destruction in 83.3%. Transfusion of type A or B blood to type AB cats results in no apparent clinical transfusion reactions.