Teratogenicity testing of BI 58 EC (38% dimethoate) in chicken embryos with special respect to degradation of the active ingredient.

The insecticide formulation BI 58 EC was tested for teratogenicity in chicken embryos, with particular reference to degradation of the active ingredient (dimethoate) after the treatment of embryonated eggs. The pesticide was diluted in water to a concentration level of 0.8%, and the emulsion was injected into the air space in a volume of 0.1 ml/egg, or hen's eggs were treated by the immersion technique. Residues of dimethoate were measured in the samples on days, 13, 15 and 19 of the incubation of chicken embryos, and morphological examinations were performed simultaneously. Analytical chemistry data indicated a slower degradation of dimethoate in embryos after the immersion of eggs, and cyllosis was remarkable in this group among the sporadic developmental anomalies. The liver tissues of both treated groups exhibited severe fatty infiltration.     

Changes in blood plasma biochemistry of chicken embryos exposed to various pesticide formulations

Wofatox 50 EC (methylparathion 50%), Nevifosz 50 EC (phosmethylan 50%), Kolfugo 25 FW (carbendazim 25%) and Dikamin D (2,4-D 40%) pesticide formulations were used as test material. The incubated chicken eggs were directly exposed to the applied pesticides with injection into the air cell. Blood samples were obtained and some plasma parameters including packed cell volume (PCV), total protein, glucose, cholesterol, aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and plasma pseudocholinesterase (PChE) activities were evaluated. Although the mortality rate obtained for the treated and control groups did not differ, there were significant changes in plasma biochemistry in relation to pesticide treatment. The present paper attempts to help those undertaking embryological and teratological studies on avian embryos exposed to pesticides including studies on changes occurring in certain plasma parameters.     

Evaluation of a modified-live virus vaccine administered in ovo to protect chickens against Newcastle disease.

The B1 strain of Newcastle disease virus (NDV-B1), which is nonpathogenic for newly hatched chickens, killed embryos when it was used to inoculate chicken eggs at embryonation day 18. Treatment of NDV-B1 with an alkylating agent, ethylmethane sulfonate (EMS) markedly reduced the pathogenicity of the virus for 18-day-old chicken embryos. Eggs inoculated with the modified virus (NDV-B1-EMS) hatched, and the virus was isolated from lungs and spleen of 1-day-old chickens. The hatched chickens developed antibody to NDV and were protected against challenge exposure (at 4 weeks of age) with a highly virulent GB-Texas strain of NDV. Presence of maternal antibody to NDV in embryonating eggs did not influence the protective ability of NDV-B1-EMS, which also induced protective immunity when administered to 4-week-old chickens. The 50% protective dose of NDV-B1-EMS in maternal antibody-negative and -positive embryos was calculated to be 10.77 and 17.70 embryo 50% lethal doses, respectively. Results of the study indicated that NDV-B1-EMS may be used as an embryo vaccine to protect chickens against Newcastle disease.     

Teratological examination of the insecticide methylparathion (Wofatox 50 EC) on pheasant embryos. 1. Morphological study

Teratogenic effects of methylparathion / Wofatox 50 EC/ and Parathion 20 WP as positive control material were tested in pheasant embryos. On the 12th day of incubation 0.1 ml of the emulsions or suspensions of the insecticides at different concentrations was inoculated into the air space of embryonated eggs. The following dose levels were employed: 27 and 270 mg/kg egg of Wofatox 50 EC, and 10 mg/kg of Parathion 20 WP. Morphological changes were evaluated by macroscopic, skeletal staining and light microscopic examinations of the embryos. Primary hypoplasia or atrophy developed in the cervical musculature /m. longus colli/ accompanied by lordosis and scoliosis of the cervical spine. In most cases we also found cyllosis .     

Embryonic toxicity of insecticide Sumithion 50 EC and herbicide Fusilade S in pheasants after individual or combined administration

The purpose of this work was to determine the individual and combined effects of insecticide Sumithion 50 EC (50% fenitrothion) and herbicide Fusilade S (12.5% fluazifop-P-butyl) on the development of pheasant embryos. Eggs were treated by injection of various concentrations of pesticides into the air space on day 12 of incubation. Pathological examination of embryos was carried out on day 23 of the hatching period. Mortality rate, body weight data and morphological alterations were evaluated after the macroscopic examination. The skeletal staining method was used to detect deformities. The two pesticides used in combination moderated the toxic/teratogenic effects of individual treatment.     

Teratological examination of the insecticide methylparathion /Wofatox 50 EC/ on pheasant embryos. 1. Morphological study

Teratogenic effects of methylparathion /Wofatox 50 EC/ and Parathion 20 WP as positive control material were tested in pheasant embryos. On the 12th day of incubation 0.1 ml of the emulsions or suspensions of the insecticides at different concentrations was inoculated into the air space of embryonated eggs. The following dose levels were employed: 27 and 270 mg/kg egg of Wofatox 50 EC, and 10 mg/kg of Parathion 20 WP.Morphological changes were evaluated by macroscopic, skeletal staining and light microscopic examinations of the embryos. Primary hypoplasia or atrophy developed in the cervical musculature /m. longus colli/ accompanied by lordosis and scoliosis of the cervical spine. In most cases we also found cyllosis.     

复州牛超数排卵效果研究

为了建立一个合理有效的复州牛超数排卵方案,获得更多更优的胚胎,长期保存复州牛的基因,试验从不同批次、哺乳情况、不同来源促卵泡素(FSH)、不同FSH剂量等方面分析了47头复州牛超数排卵效果.结果表明,经超数排卵处理的复州牛头均回收卵数7.98枚,其中头均有效胚数4.62枚;供体牛体况好、超数排卵方案细化的第3批次效果最好,头均有效胚数8.25枚,显著高于第1批次(2.00枚)和第2批次(2.79枚)(P <0.05);未哺乳牛头均黄体数、头均回收卵数和头均有效胚数分别为15.38个、7.63枚、5.75枚,极显著高于哺乳牛的10.64个、2.18枚、0.64枚(P <0.01);日本产FSH头均回收卵数、头均有效胚数分别为12.70和7.50枚,动物所产FSH分别为10.62和7.38枚,二者间差异不显著(P >0.05);动物所产FSH的适宜剂量为每头8.8 mg,头均有效胚数7.38枚,有效胚率为69%.通过优化复州牛的超数排卵方案,取得了理想的超数排卵效果.     

盐酸刺激量对家蚕卵酯酶A4活性的影响及其与蚕卵活化的关系

将产卵后 2 4～ 96h( 2 5℃ )的蚕卵 ,用相同或最佳盐酸刺激量 (浸酸后点青最快、点青卵率最高 )浸酸处理 ,卵龄越大 ,蚕卵点青越迟。蚕卵的点青时间在一定范围内随盐酸刺激量增大而提前 ,超过这一范围 ,则随刺激量增大而延迟。卵龄增大 ,蚕卵酯酶A4和ATPase特征性活性峰出现时间延迟 ;蚕卵经盐酸处理 ,该时间明显提前。产卵后 96h盐酸处理蚕卵 ,酯酶A4的活性峰出现时间随盐酸刺激量增大而提前。随着浸酸卵龄和盐酸刺激量的改变 ,蚕卵酯酶A4的活性峰出现时间与蚕卵点青时间发生一致的变化。盐酸刺激量能够影响蚕卵的活化 (或滞育发育期 )和发育速度。酯酶A4的ATPase活性变化能够反映盐酸处理蚕卵的滞育程度及所使用的盐酸刺激量     

不同FSH对和牛超数排卵效果的影响

[目的]以16~18月龄青年日本和牛为供体,研究不同FSH和剂量对青年日本和牛超排效果的影响。[方法]超排采用CIDR+FSH+PG方法进行。[结果]进口FSH和国产FSH超排和牛后头均获胚胎数、头均获可用胚胎数、头均退化胚胎数和头均未受精卵数差异均不显著(P0.05);3种不同剂量的国产FSH处理日本和牛,头均获胚胎数、头均获可用胚胎数、头均退化胚胎数和头均未受精卵数差异均不显著(P0.05)。[结论]进口FSH和国产FSH超排青年和牛效果相当,同时适当降低国产FSH注射总剂量不影响青年和牛供体超排效果。     

Clinical epidemiologic and experimental evidence for the transmission of Newcastle disease virus through eggs

Sporadic outbreaks of Newcastle disease (ND) occurred in Taiwan during 1998-2000. In some cases, the disease occurred in broilers less than 2 wk old that originated in a broiler breeder farm, so spread of the ND virus (NDV) from the infected breeder farm to broiler ranches was suspected. The purpose of the present study was to examine the possibility of the transmission of NDV through eggs. Both clinical and experimental evidence were used to prove that this is possible. From epidemiological investigation, the possibility of transmission through eggs was suggested in two separate ND cases from a breeder farm and its progeny because two identical NDVs were isolated from both cases. In order to clarify the possibility of the transmission through eggs, one mean egg lethal dose (ELD50) of NDV was inoculated into the allantoic cavity of 155 9-to-11-day-old specific-pathogen-free (SPF) chicken embryos. Seventy-one hatching chicks from the inoculated embryos were raised for 14 days. The cloacal swabs from those chicks at the ages of 1, 4, and 7 days and the tissues after necropsy at the ages of 14 days were taken for virus isolation. The same NDV was reisolated from three hatching chicks. This experiment confirms that a few chicken embryos infected in ovo with a low titer of NDV can hatch and contain NDV after hatching, which results in NDV spreading through eggs.     

Descent of ova and embryos in cows superovulated with serum gonadotropin]

The descent and localization of eggs and embryos in individual segments of the reproductive tract of superovulated cows were studied in this work. For the induction of superovulation, serum gonadotropin (PMSG, Ivanovice in Haná) at a dose of 2,500-3,500 I.U. was used, in combination with 0.5 mg of Cloprostenol (Oestrophan, Spofa), administered 48 hours after gonadotropin treatment. The start of superovulation fell on days 9 to 12 of the sexual cycle and was conditioned by the presence of the corpus luteum (CL). After the onset of the heat, 2-3 inseminations were carried out using fresh semen. Donor cows were slaughtered 3, 4, 5, 6, and 7 days after the second insemination and isolated reproductive organs (Fig. 1) were divided into five segments (two on oviducts and three on uterine horns) by the applied ligature. In laboratory conditions superovulation response was determined accurately, the volume of ovaries was assessed according to water displacement and the segments of oviducts and uterus were rinsed with TCM 199 or PBS supplemented with FCS. 3, 4, 5, 6, and 7 days after insemination (Tab. I). 18.1 (+/- 3.55), 12.4 (+/- 0.91), 19.2 (+/- 2.86), 20 and 23 (+/- 2.44) CL on average were recorded, which corresponded to the ovulation of 64, 50, 56, 71 and 72 percent of stimulated follicles (Fig. 2). Within 3 to 7 days after insemination nearly triple enlargement of ovaries was also observed (Tab. I, Fig. 3). During the lavage of individual segments of the tubular reproductive tract, 38 per cent of eggs and embryos were detected in the uterus as early as 3 days after insemination (Tab. II). Unfertilized eggs and degenerated embryos were found in the 2nd and 3rd uterine segment, embryos at the stage of 8-16 blastomeres were localized in the 1st and 2nd segment of the uterus. Four days after insemination (Tab. III), about 64 per cent of eggs and embryos at the stage up to 16 blastomeres were found in the uterus, but embryos up to 32 blastomeres were still flushed out of the oviduct. On day 5 after insemination, 92 per cent of eggs and embryos were released into the uterus, being localized mostly in the cranial and medial part of the uterus (Tab. IV). 7.5 per cent of recovered eggs and embryos at the stage of early or compacted morulae were still detected in the oviducts.(ABSTRACT TRUNCATED AT 400 WORDS)     

Plasma inhibin A determination at start superovulatory FSH treatments is predictive for embryo outcome in goats

To test whether inhibin A assays can be used for the prediction of yields in embryo programmes in goats, 50 does were treated with 45 mg FGA sponges (Chronogest) for 16 days plus a single dose of 100 microg i.m. cloprostenol on Day 14, just before the start of administration of eight doses of 1.25 ml of Ovagen twice daily for 4 days. At first FSH injection, the number and size of all follicles > or =2 mm was assessed by transrectal ultrasound and plasma inhibin A levels were measured by specific dimeric assay. There was a positive correlation between number of follicles > or =6 mm (8.8 +/- 0.5) and inhibin A levels at first FSH dose (193.2 +/- 14.5 pg/ml, P<0.05). The mean number of corpora lutea on Day 7 after sponge removal was related to the total number of follicles with a diameter of 2-6 mm at the onset of the FSH treatment (15.3 +/- 0.7, P<0.05). The total number of embryos recovered was related to the number of follicles with 4-6 mm in size (6.2 +/- 0.5, P<0.05) and to the inhibin A levels at first FSH dose (P<0.05). These results suggest that follicles > or =4 mm are the source of inhibin prior to FSH stimulation and are the main source of oocytes resulting in the number of viable embryos recovered after a superovulatory treatment. Hence, the response to superovulatory treatments in goats in terms of the number of embryos can be predicted from either the population of follicles determined by ultrasound or the plasma inhibin A levels at start of the superovulatory FSH treatment.     

Survivability of Infectious Hematopoietic Necrosis Virus in Fertilized Eggs of Masu and Chum Salmon

Abstract

The possibility of vertical transmission of infectious hematopoietic necrosis virus (IHNV) was studied with the eggs of masu (cherry) salmon Oncorhynchus masou and chum salmon O. keta. The surfaces of eggs and sperm were contaminated with IHNV (10^3.8-10^4.8 50% tissue culture infective dose [TCID50]/egg) and then the eggs were fertilized. Eggs just after fertilization and embryonated eggs also were infected by injection with IHNV (10^3.8 TCID50/egg) directly into the yolk. During incubation, eggs were held in running water at 10°C. Mortality of the eggs or hatched progeny was determined and isolation of IHNV on the surface or inside of the eggs was determined during the incubation period. No mortality occurred and no virus was detected in fertile eggs from contaminated gametes. For injected eggs, IHNV was not detected on the surface of masu and chum salmon eggs after 1 d of incubation. Infectivity of IHNV inside the eggs decreased gradually and could not be detected after 1 month of incubation. This rate of IHNV reduction in the fertilized egg was similar to that found in a mixture of IHNV and homogenized yolk contents. Several individual yolk components also showed anti-IHNV activity. When eyed eggs were injected with IHNV, the embryos of both masu and chum salmon became infected, and the concentration of virus increased rapidly and reached more than 10^6.5 TCID50/fish. The cumulative mortality of eggs injected at the eyed stage for both masu and chum salmon was 90%. The susceptibilities of hatched-out larvae of masu and chum salmon to IHNV were different; cumulative mortality was more than 90% in masu salmon and 20–30% in chum salmon artificially infected with the virus. We concluded that vertical transmission of IHNV is doubtful because the virus is apparently unable to survive in eggs before the eyed stage.     

Degradation of some pesticides in avian embryos

On day 9 or 12 of the hatching period different pesticides (parathion, methyl-parathion, carbendazim, 2,4-D-amine Na, phosmethylane) were applied in ecotoxicological trials. The formulations were either injected into the air space of pheasant, quail or hen eggs or hen eggs were treated by the immersion technique. The residues of pesticides were measured in samples on days 13, 14 and 16 of incubation of chicken and pheasant embryos, while the Japanese quail embryos were analysed on days 10-14 of incubation. Analytical chemistry data showed a varying degradation rate of the compounds in avian embryos of the same species. The residues directly affect the embryos, disturbing their normal development and causing pathophysiological and morphological changes.     

Effect of cadmium injected in ovo on hatching results and the activity of plasma hydrolytic enzymes in newly hatched chicks

The aim of the study was to determine the toxicity of cadmium ions in chick embryos, using plasma hydrolytic enzyme as its biomarker. Hatching eggs (n = 300) from Ross 308 broilers were incubated under standard conditions. On day 4 of incubation, 50 μl of saline solution, containing Cd ions at a concentration from 0 (control group) to 24 μg, was injected in ovo into the egg albumen. The results indicate that the administration of cadmium at doses exceeding 1 μg/egg caused a gradual decrease in hatchability, with an LD50 of 3.9 μg/egg. The greatest differences between the groups in the enzymatic activities studied were found for N-acetyl-β-D-glucosaminidase (NAG), β-D-mannosidase (β-MAN) and arylsulphatase (ARYL). Compared to the control group, in the blood serum of chicks from the groups receiving 3, 6 and 12 μg Cd/egg the NAG activity increased by 79, 108 and 54% and β-MAN activity by 33, 119 and 108%, respectively. Exposure to cadmium at a dose of 1 to 6 μg per egg caused an about 60% increase in ARYL activity while a dose of 12 μg decreased the activity by about 35% below the level observed in the control group. These findings show that cadmium has a similar toxicity mechanism in mammals and birds, which opens the possibility of using NAG activity as a biomarker of the cytotoxic effect of cadmium in birds.     

Pathological and immunological study of goose embryos and goslings inoculated with an attenuated strain of Derzsy's disease virus.

An attenuated Derzsy's disease virus strain, designated BAV, was studied in goose embryos. A total of 248 embryonated goose eggs, coming from a susceptible laying flock with no yolk-derived immunity (group I) and from a vaccinated laying flock (group II) were used. The eggs were inoculated into the allantoic cavity with 10(1.9), 10(2.9) or 10(3.9) EID 50/0.2 ml virus on day 12 or day 20 of incubation. Embryos were killed at 5-day intervals. The dead embryos and the hatched goslings (up to 2 weeks of age) were examined by gross and histopathological methods. Reisolation of the virus from the organs was attempted, serum samples were tested for the presence of antibodies, and lymphocytes separated from the circulating blood were used in the lymphocyte stimulation and immunorosette formation tests. Embryos of both groups I and II, inoculated at either time of incubation, showed a body mass gain inferior to that of the controls. Sixteen (group I) and 12 (group II) of the embryos inoculated on day 12 of incubation died. Some (group I: 15, group II: 6) of the embryos inoculated on day 20 of incubation failed to hatch. The pathomorphological changes seen in the embryos killed between days 17 and 22 of incubation were of degenerative character. In embryos killed later (between days 23 and 58 of incubation) the degenerative changes were accompanied by infiltration by inflammatory cells. Reisolation of the virus strain was mostly successful between postinoculation (PI) days 5 and 10. Specific virus-neutralizing antibodies and cellular immune response were demonstrable already at hatching.(ABSTRACT TRUNCATED AT 250 WORDS)     

Development of porcine embryos in vitro: effects of culture medium and donor age.

We compared development of porcine embryos in three media and evaluated the effect of age of the donor on embryo development in vitro. In Exp. 1, embryos were collected from 35 postpubertal females on d 2 or 3 after onset of estrus. Embryos were cultured 144 h in Whitten's Medium (WM), North Carolina State University Medium-23 (NCSU-23), or Beltsville Embryo Culture Medium-3 (BECM-3) in 95% air: 5% CO2 at 39 degrees C. More (P < 0.01) embryos that were initially one cell or two cells developed to blastocysts when cultured in NCSU-23 (56%) and BECM-3 (43%) rather than in WM (7.5%). More (P < 0.01) embryos that were four cells at recovery developed to blastocysts in NCSU-23 (97%) than in BECM-3 (69%) or WM (69%). Blastocysts that developed from four-cell embryos cultured in BECM-3 had more (P < 0.01) nuclei than blastocysts that developed from four-cell embryos in the other two media. In Exp. 2, ovarian responses, fertilization rates, and in vitro embryo development in NCSU-23 and BECM-3 were compared for postpubertal (approximately 170-d-old) gilts vs gilts given exogenous gonadotropins at 102 d of age. Ovulation rate (P < 0.01), number of eggs recovered, and number of eggs fertilized per gilt (P < 0.001) were greater in the older gilts. The percentage of eggs fertilized, the number of unfertilized eggs, and the number of unclassifiable eggs were similar (P > 0.10) for both age groups. More (P < 0.10) blastocysts developed from embryos recovered from 170-d-old than from 102-d-old gilts, and more (P < 0.05) blastocysts developed in NCSU-23 than in BECM-3. Zona thicknesses and number of nuclei per embryo were similar (P > 0.10) for both ages. We conclude that embryos from prepubertal gilts do not have the same in vitro developmental potential as those from cyclic gilts. However, superior development of embryos in NCSU-23 from both 102-d-old and 170-d-old gilts indicates that media composition did not differentially affect embryos produced by younger vs older gilts.     

家蚕的半活化胚现象初探

滞育是家蚕的生理现象 ,家蚕滞育卵的活化处理是养蚕生产上重要的技术操作。研究了家蚕盐酸处理卵、复式冷藏卵和长期保存卵的胚胎活化过程 ,认为家蚕胚胎除滞育态和活化态以外 ,还存在半活化状态。半活化胚胎的形态与萌动期 (乙 1)至反转期 (己 1)的活化胚相似 ,但胚胎整体发育不平衡 ,后部发育慢于前部 ,在 2 5℃、RH 75 %～ 85 %催青环境中的发育速度明显慢于活化胚 ,而越接近反转期发育速度越接近活化胚。同一群体内出现的半活化胚胎之间发育比较整齐 ,而不同群体的半活化胚胎之间发育有一定差异。活化刺激量越大 ,出现的半活化胚胎发育越快。探讨了半活化胚胎产生的机理以及对蚕种生产的影响     

Growth kinetic studies of avian infectious bronchitis virus in tracheal organ cultures.

An egg-adapted vaccine strain (H120) and an organ culture-passaged field strain (HV-10) of avian infectious bronchitis (AIB) virus were propagated in tracheal organ cultures and their growth kinetics examined using nine-day-old embryonated fowl eggs and chick tracheal explants for virus assay. When the H120 strain was assayed in embryonated eggs, titres were approximately log10 2-0 ID50 (50 per cent infectious dose) per ml higher than when assays were performed in tracheal explants. The HV-10 strain, assayed in tracheal explants, yielded higher titres than did the H120 strain, but when assayed in embryonated eggs, yielded only minimal and variable virus titres.     

Effects of short-term exposure to condensed tannins on adult Trichostrongylus colubriformis

Twelve parasite-naive sheep were used to study the possible direct anthelmintic effect of a condensed tannin extract (quebracho) on the population and fecundity of the intestinal nematode Trichostrongylus colubriformis. The sheep were infected with a single dose of 20,000 L3 of T colubriformis. Twenty-eight days later, six of them were drenched daily for a week with quebracho extract at 8 per cent by weight of their food intake. All lambs were then slaughtered, and their small intestines removed to estimate the worm burdens and the numbers of eggs in utero. Two days after the first drench with tannin extract the faecal egg counts of the treated sheep were approximately 50 per cent of those of the control sheep (P<0.01), but there was no further reduction with continued drenching. In the treated sheep the worm burdens and number of eggs per gram faeces per worm were reduced by 30 per cent compared with the controls (P<0.05), but the sex ratios, the number of eggs in utero and length of the worms were not affected by drenching with tannin.