Effect of Follicle Size on In Vitro Maturation of Pre‐Pubertal Porcine Cumulus Oocyte Complexes

Very small follicles (<3.0 mm diameter) are over‐represented on the surface of ovaries of non‐cycling pigs, and the oocytes collected from these follicles generally have reduced developmental competence in vitro. This study examined the effect of follicle size on the nuclear maturation (n = 608), the potential of parthenogenetic activation (n = 243) and the cyclic AMP (cAMP) content of pre‐pubertal porcine oocytes (n = 480). In addition, the influence of follicle size on steroid hormone synthesis was analysed. Cumulus oocyte complexes (COCs) flushed from small (2.5–4.0 mm) or large (4.5–6.0 mm) ovarian follicles were cultured for 0, 28 and 46 h. After 46 h of IVM, a greater proportion of oocytes from 4.5‐ to 6.0‐mm follicles reach metaphase II (MII) compared with those from follicles with 2.5–4.0 mm of diameter (96.1 vs 77.0%, respectively; p < 0.001). Parthenogenetic activation of oocytes from large follicles produced higher developmental rates than oocytes from large follicles (p < 0.05). At 28 h, the IVM medium with oocytes from large follicles contained significantly more 17ß‐oestradiol (E₂) than the medium with oocytes from small follicles (5.55 vs 3.45 ng/ml, respectively; p < 0.05) and at 46 h, the medium with oocytes from small follicles contained significantly more progesterone (P₄) than the medium with oocytes from large follicles (276.7 vs 108.2 ng/ml, respectively, p < 0.05). Porcine oocytes from large follicles have higher nuclear and cytoplasmic maturation capacities, but the differences did not appear to be cAMP‐mediated. Our findings also suggest that COCs from small follicles undergo more intensive luteinization than COCs from large follicles. The results show that oocytes from follicles with a diameter greater than 4.0 mm are more suitable for in vitro studies.     

Direct desaturation of free myristic acid by hen liver microsomal Delta9-desaturase without prior activation to myristoyl-CoA derivative

Direct desaturation of free myristic acid by hen liver microsomal Delta(9)-desaturase without prior activation to myristoyl-CoA by the addition of adenosine triphosphate (ATP) and CoA was observed when the incubation medium was mixed at mixing speeds of >250 rpm in the presence of fatty acid-binding proteins (FABP). Desaturation was linear with time and proportional to the microsomal protein concentration. Desaturation was maximal at pH 7.9. The greatest desaturation rate was observed at a mixing speed of 500 rpm in the presence of FABP. Desaturation decreased at mixing speeds of >500 rpm. Data suggest that when myristic acid is bound to FABP in the form of protein-monomer complexes, its activation to the CoA derivative is not necessary for it to be desaturated by the Delta(9)-desaturase when using mixing rates of >250 rpm. Myristic acid-FABP complexes serve as substrates for the Delta(9)-desaturase at mixing rates of >250 rpm. Desaturation was reduced by bovine serum albumin and alpha-bromohexadecanoate, and no desaturation was observed in the absence of FABP. These findings suggest that FABP may regulate the accessibility of fatty acids in the desaturation reaction to the active site of the desaturase rather than just protect the membrane-bound desaturase from the cytotoxic effect of free fatty acids.     

Using indirect ELISA to assess different antigens for the serodiagnosis of Fasciola gigantica infection in cattle, sheep and donkeys

An indirect enzyme-linked immunosorbent assay (iELISA) was evaluated for its diagnostic capability in detecting antibodies against Fasciola gigantica infection in cattle, sheep and donkeys sera using crude worm, excretory–secretory and glutathione S-transferase antigens prepared from adult liver fluke. Presence of F. gigantica worms at post-mortem examination of cattle, sheep and donkey’s livers was taken as a gold standard for the evaluation of the assay. The diagnostic sensitivity, specificity and accuracy percentages of iELISA were determined for each antigen. Excretory–secretory antigen gave the best results for the serodiagnosis of F. gigantica infection in cattle, sheep and donkeys using iELISA with diagnostic sensitivity percentages of 93.3%, 94.9% and 93.3%, respectively, while the specificity percentages were 96.7%, 97.2% and 96.3%, respectively, whereas the accuracy percentages were 95%, 96% and 95.7%, respectively. The diagnostic sensitivity percentages of iELISA using crude worm antigen were 96.7%, 100% and 93.3%, respectively, while the specificity percentages were 80%, 83.3% and 85.2%, respectively, whereas the accuracy percentages were 88.3%, 86.7% and 87%, respectively. The diagnostic sensitivity percentages of iELISA using glutathione S-transferase antigen were 66.7%, 71.8% and 60%, respectively, while the specificity percentages were 70%, 77.8% and 77.8%, respectively, whereas the accuracy percentages were 68.3%, 74.7% and 73.9%, respectively. Conclusively, excretory–secretory antigen dependent iELISA can be used as a reliable serodiagnostic test for F. gigantica infection in cattle, sheep and donkeys.     

Reproductive Performance of Gilts with Similar Age but with Different Growth Rate at the Onset of Puberty Stimulation

The aim of this study was to evaluate the reproductive performance of gilts that had a similar age but different weights at the onset of puberty stimulation by boar exposure at 144 days. Gilts were divided into two groups according to their lifetime growth rate from birth to approximately 144 days of age. Mean growth rates at this moment were 577 and 724 g/day for group 1 (G1; n = 58) and group 2 (G2; n = 58), respectively. After selection, gilts were weighed at approximately 155, 165 and 175 days of age, on the insemination day and at slaughter. Gilts were inseminated, on average, at 193 days of age and were slaughtered 32 days after insemination, when the number of corpora lutea and embryos were recorded. Higher growth rate gilts (G2) reached puberty earlier (155.3 vs 164.1 days; p < 0.01). More gilts of G2 group attained puberty by 190 days of age (p = 0.004) than G1 gilts (95%; 55/58 vs 76%; 44/58). The anoestrous rate, until 60 days after the onset of boar exposure was higher (p < 0.01) in G1 (19.0%; 11/58) than in G2 (3.4%; 2/58) group. However, there were no differences in the pregnancy rate (90.7 vs 94.5), ovulation rate (15.9 vs 16.5), total embryos (12.9 vs 11.7), viable embryos (12.0 vs 11.1) and embryo survival (73.7% vs 68.5%), between G1 gilts and G2 gilts, respectively (p > 0.05). High growth rate gilts attain puberty earlier and have a lower anoestrous rate than low growth rate gilts.     

Minimal External Masculinization in a SRY-negative XX Male Podenco Dog

Normal mammalian sex differentiation takes place in three genetically controlled steps: chromosomal sex determination (XX or XY), gonadal differentiation and development of the phenotypic sex. Animals are considered to be sex reversed if chromosomal sex determination and gonadal development are not in agreement. In this report, sex reversal is described in a 1.5-year-old Podenco dog that was referred because of suspected recurrent growth of a previously removed os clitoridis in the vulva. With that exception the dog was phenotypically female, but had never been in oestrus and exhibited male behaviour. Abdominal ultrasonography showed a small tubular structure dorsal to the bladder, consistent with a uterus. An ovoid structure resembling a gonad was visible between the right kidney and inguinal canal. Plasma testosterone concentrations before and after GnRH administration indicated the presence of functional testicular tissue. Two testes, each with its epididymis and ductus deferens, and a complete bicornuate uterus were removed surgically. Cytogenetic analysis of peripheral blood lymphocytes showed a normal female karyotype (78, XX). These findings are consistent with the diagnosis of an XX male. PCR analysis of genomic DNA revealed that the SRY gene was absent. In summary, this report describes the first SRY-negative XX male Podenco dog with an almost complete female phenotype despite high basal and stimulated plasma testosterone concentrations. It is hypothesized that the clinical observations in this dog may have been caused by reduced and delayed Müllerian-inhibiting substance secretion and the absence of conversion of testosterone to dihydrotestosterone due to 5α-reductase deficiency.     

Isolation, propagation, identification and comparative pathogenicity of five Egyptian field strains of Eimeria tenella from broiler chickens in five different provinces in Egypt

A battery trial was conducted to evaluate the comparative pathogenicity of five field strains of Eimeria tenella from Behera, Khafr El-Sheikh, Alexandria, Gharbia and Matrouh provinces in Egypt. Two-week-old broiler chickens were infected with 25×10(3) sporulated oocysts of each strain of E. tenella. The comparative pathogenicity of the strains was assessed by calculating body weight gain, feed conversion ratio, lesion scores, dropping nature scores, cecal scrapings, mortality percentage and oocysts count. Hematological parameters including hemoglobin (Hb) content, packed cell volume (PCV%) and total erythrocytic count, were also evaluated. There were different degrees of pathogenicity between the strains.     

Epidemiology of tomato yellow leaf curl begomovirus in the Fayium area, Egypt

Tomato yellow leaf curl begomovirus (TYLCV) severely invaded tomato plantations in Egypt (Lower and Middle Egypt) in 1989. This study aimed to discover the relationship between TYLCV and other epidemic-associated factors in the Fayium area. The rate of TYLCV infection was inspected visually for three successive years (1994/1996) in the Fayium area. During the same period, whiteflies were collected for virus detection using bait-plant and DNA hybridization techniques. DAS-ELISA was used to detect mixed virus infections in tomato plants. TYLCV infection was prevalent (60–68%) and severe (2.1–3.0) in the Fayium fields. Cucumber mosaic cucumovirus (CMV) was found in some fields (5–28%) with moderate severity (1.0–20). Potato Y potyvirus (PVY) and potato leaf roll polerovirus (PLRV) were found in few fields (5–19% and 5% respectively) at very low severity. There was a negative correlation between TYLCV occurrence and distance from the source of infection, and a positive correlation (98%) between TYLCV intensity and percentage of viruliferous whiteflies in 1994 and 1995. There was no positive correlation between TYLCV and the total population of whiteflies caught during the same period. Five percent of viruliferous whiteflies, as proved by cDNA hybridization, led to 46% TYLCV infection. The same percentage of whiteflies, as shown by bioassay, led to 68% TYLCV infection. Monitoring of viruliferous whiteflies could be used for early prediction of TYLCV infection.     

Differences in Antennal Sensillae of Male and Female Peach Fruit Flies in Relation to Hosts

Antennal sensillae of male and female peach fruit flies, Bactrocera zonata (Saunders) (Diptera: Tephritidae), obtained from three different host fruit species (guava, Psidium guajava L. (Myrtales: Myrtaceae); peach, Prunus persica (L.) Stokes (Rosales: Rosaceae); and orange, Citrus sinensis (L.) Osbeck (Sapindales: Rutaceae)), were studied with scanning electron microscopy. This study was carried out to describe the different types of sensillae present on the three antennal segments (scape, pedicel, and flagellum or funiculus) of both sexes of B. zonata on different host fruit. The antennal segments of females tended to be larger than those of males feeding on peach and guava fruit. On orange, both sexes were similar (no significant differences were found). The first two antennal segments, scape and pedicel, are reinforced by some bristles and have different types of sensillae, including trichoid I, II, S; basiconic II; and sensilla chaetica in different numbers on different host fruit species. Numerous microtrichia, as well as trichoid (I, II), basiconic (I), clavate, and coeloconic (I, II) sensillae were observed on the funiculus with a great variation in number and length. As a result of feeding on different hosts, differences were found between sexes and some plasticity in size, number, distribution, and position of some sensillae, including trichoid, basiconic, chaetica, and clavate on the antennae of the female B. zonata. These sensillae were significantly larger in females. Also, some morphological and morphemetric differences have been found according to their feeding on different host fruit.     

Reproductive Development of Santa Inês Rams During the First Year of Life: Body and Testis Growth,Testosterone Concentrations,Sperm Parameters,Age at Puberty and Seminal Plasma Proteins

We have investigated the reproductive development of the tropically adapted Santa Inês ram, the most common hair sheep in Brazil. From 8 to 48 weeks of age, 16 animals were evaluated for body and testis growth, semen parameters, testosterone concentrations and seminal plasma proteins, using two‐dimensional SDS‐PAGE. Animals were weaned at 30 days and kept in feedlots thereafter, receiving hay, concentrate (18% of crude protein) and mineral supplement. Body weight increased from 12.3 ± 0.7 to 54.3 ± 1.6 kg between 8 and 48 weeks (p < 0.05), but changes in thoracic perimeter and scrotal circumference were non‐significant after 36 weeks (p > 0.05). The percentage of motile sperm increased slowly until 23 weeks and more rapidly after that age, but significant changes in progressive motility occurred after 25 weeks. Presence of abnormal sperm related inversely to age. Most significant changes in sperm concentration occurred between 38 and 44 weeks (0.38 ± 0.05 to 1.14 ± 0.24 × 10⁹ cells/ml, p < 0.05) and testosterone reached its highest concentrations at 42 weeks, decreasing afterwards. Rams reached puberty at 28.2 ± 0.8 weeks. The number of protein spots on seminal plasma gels was similar from 15 to 18 weeks (45 and 47 spots; p > 0.05), increased until 24 weeks (141 spots) and 28 weeks (170 spots; p < 0.05) and remained without significant (p > 0.05) changes from 28 to 48 weeks (186 ± 10 spots). Furthermore, the intensity of selected spots on 2D maps increased (p < 0.05) between 15 and 28 weeks, which preceded or coincided with the main developmental changes in sperm motility and percentage of defective sperm in the ejaculates. These results will support future studies designed to characterize specific seminal plasma proteins whose expression relate to the development of testis, epididymis and accessory sex glands.