Induction of tibial dyschondroplasia by carbamate and thiocarbamate pesticides

Tibial dyschondroplasia (TD) is a major poultry leg problem, the natural etiology of which is unknown. Certain dithiocarbamate pesticides such as tetramethyl thiuram disulfide (thiram) have been shown to induce the disease in chickens. Because many different carbamate and thiocarbamate chemicals are used in a number of agricultural, industrial, and household applications, the objective of this study was to determine whether all chemicals of these categories induce TD and whether there is a concentration-dependent relationship between the ingestion of these chemicals and the incidences and the severity of the disease. Week-old broiler chicks were fed diets containing thiram or other assorted carbamate and thiocarbamate pesticides mixed in feed for 24-48 hr between ages 8 and 10 days. The birds were killed on day 15 and the proximal tibial and tarsometatarsal growth plates were evaluated for the presence and severity of TD lesions. TD was distinguished by broadening of growth plates; upon histologic exam chondrocytes appeared to be shrunken and dead. When compared by including equimolar concentrations of these chemicals in the feed, the dithiocarbamates with more than two sulfide groups, such as disulfiram, ferbam, thiram, and ziram were potent inducers of TD, whereas those with two sulfides to no sulfide group appeared ineffective at inducing TD. Both thiram and ferbam also reduced the bird's body weights. Thiram increased the incidence and the severity of the disease, denoted by TD index, in a dose-dependent manner. These results suggest that inadvertent contamination of feed or litter with some of these or similar chemicals may cause leg problems in poultry.     

In vitro production of sexed embryos for gender preselection: high-speed sorting of X-chromosome-bearing sperm to produce pigs after embryo transfer

The objectives for the present experiments were to apply sperm sexing technology to an in vitro production system with porcine oocytes obtained from slaughterhouse material. On six experimental days, ovaries were obtained from an abattoir, and cumulus-oocyte-complexes were matured in vitro. Semen was collected from mature boars of proven fertility and was sorted for X-chromosome-bearing sperm, using the Beltsville Sperm Sexing Technology incorporating the use of high-speed sorting. A total of 5,378 oocytes were submitted for in vitro fertilization (IVF). Of these, 559 ova were stained for cytogenetic analysis 18 h after IVF. From the remaining 4,819 ova, 1,595 cleaved, and 1,300 of the cleaved embryos were transferred into 26 synchronized recipients (5 control gilts for unsorted sperm, 21 gilts for X-sorted sperm). In a test of two fertilization media (FERT-A vs FERT-B) higher cleavage rates (P<.05) were obtained when FERT-B was used as a fertilization medium for unsorted (43.4+/-5.1%) and sorted sperm (43.1+/-1.1%;), whereas in FERT-A unsorted sperm gave a cleavage rate of 17.9+/-4.4% and sorted sperm gave 30.4+/-1.4%. Additionally, cleavage rates were higher (P<.05) after fertilization with sorted sperm vs unsorted sperm, independent of fertilization medium. Cytogenetic analysis of ova revealed that more oocytes with unsorted than with sorted sperm remained in Metaphase 2 arrest (P<.05). This was also independent of the fertilization medium. Monospermic fertilization rates were the same for IVF with unsorted or sorted sperm, independent of the fertilization system, except FERT-A with unsorted sperm (P<.05). Polyspermic fertilization rates were highest in FERT-B (37.6+/-6.6). A total of 57 pigs were born from nine litters. Six litters from sexed sperm (X-sorted) produced 33 females (97%) and one male. Three litters from control transfers produced 23 pigs, 11 of which were female (48%). The sex ratio of the offspring was predicted based on the sort reanalysis of the sorted sperm for DNA content.     

Respiratory infection of turkeys with Listeria monocytogenes Scott A

The pathogenesis of L. monocytogenes strain Scott A was studied by challenging day-old male turkey poults by air sac inoculation with tryptose phosphate broth containing 10(0) cfu (control), 10(4), 10(5), and 10(6) cfu (low challenge), or 10(7) and 10(8) cfu (high challenge) of the Scott A (serotype 4b) strain of L. monocytogenes. Mortality at 2 wk postinfection (PI) ranged from 25% for low challenge to 100% for high challenge (P= 0.0001). Gross and histopathological lesions were observed in heart, liver, spleen, lung, and bursa of Fabricius of mortalities at 4 days PI. Listeria monocytogenes challenge resulted in significantly decreased relative weight of the bursa of Fabricius and increased relative weight of the spleen, and L. monocytogenes was isolated by direct plating of liver, pericardium, brain, and both left and right stifle joint synovium (knee) cultures, as well as gall bladder, yolk sac, and cecal tonsil from transfer swabs onto Listeria-selective agar. Isolates were confirmed as positive using Gram stain, biochemical tests, and the Biolog system. High challenge resulted in confirmed L. monocytogenes isolation from 48% of left knee and 59% of right knee cultures. Low challenge resulted in isolation of L. monocytogenes from 11% of both left and right knee cultures. These results suggest that L. monocytogenes Scott A colonization of turkey knee synovial tissue can initiate in day-of-age poults and that L. monocytogenes Scott A can be invasive through air sac infection.     

Effect of liner design,pulsator setting,and vacuum level on bovine teat tissue changes and milking characteristics as measured by ultrasonography

: Friesian-type dairy cows were milked with different machine settings to determine the effect of these settings on teat tissue reaction and on milking characteristics. Three teat-cup liner designs were used with varying upper barrel dimensions (wide-bore WB = 31.6 mm; narrow-bore NB = 21.0 mm; narrow-bore NB1 = 25.0 mm). These liners were tested with alternate and simultaneous pulsation patterns, pulsator ratios (60:40 and 67:33) and three system vacuum levels (40, 44 and 50 kPa). Teat tissue was measured using ultrasonography, before milking and directly after milking. The measurements recorded were teat canal length (TCL), teat diameter (TD), cistern diameter (CD) and teat wall thickness (TWT).Teat tissue changes were similar with a system vacuum level of either 50 kPa (mid-level) or 40 kPa (low-level). Widening the liner upper barrel bore dimension from 21.0 mm (P < 0.01) or 25.0 mm (P < 0.001) to 31.6 mm increased the magnitude of changes in TD and TWT after machine milking. Milk yield per cow was significantly (P < 0.05) higher and cluster-on time was reduced (P < 0.01) with the WB cluster as compared to the NB1 cluster. Minimum changes in teat tissue parameters were achieved with system vacuum level of 40 kPa and 50 kPa using NB and WB clusters, respectively. Similar changes in teat tissue and milk yield per cow were observed with alternate and simultaneous pulsation patterns. Widening pulsator ratio from 60:40 to 67:33 did not have negative effects on changes in teat tissue and had a positive effect on milk yield and milking time. Milk liner design had a bigger effect on teat tissue changes and milking characteristics than pulsation settings.     

In vitro fertilisation of in vivo matured porcine oocytes obtained from prepuberal gilts at different time intervals after hCG injection

The goal of the present study was to find out the best interval after hCG injection in PMSG primed prepuberal gilts for retrieval of in vivo matured oocytes for in vitro fertilisation (IVF). Altogether 66 gilts were superovulated with 1500 IU PMSG and 500 IU hCG 72 h later. Ovum pick up was performed endoscopically 24, 28, 32 or 36 h after hCG and a total of 869 cumulus-oocyte-complexes (COCs) were aspirated from 1400 follicles. COCs were tested for quality, and an aliquot was immediately fixed and stained to determine meiotic configuration. The remaining COCs were fertilised in vitro using frozen-thawed epididymal semen. Quality and developmental stage of embryos were tested after IVF, and the number of nuclei was counted. At 24 to 32 h after hCG only few oocytes have entered the second meiotic cycle (18 to 25% vs. 58% at 36 h, p < 0.05). The overall cleavage rate was significantly influenced by insufficient maturation rate at the early collection times (14% at 24 h vs. 49% at 36 h). Additionally, when oocytes were collected 24 to 32 h vs. 36 h the cleavage rate based on mature oocytes was lower (26 vs. 62%, p < 0.05). Once embryonic development has been initiated, the further in vitro development to blastocyst stages did not differ between groups. However, the number of cells was lower at collection times 24 to 32 h as compared to 36 h after hCG (12 to 15 cells vs. 22 cells, p < 0.05). The results indicate that the time of COC collection affects the in vitro developmental competence up to the blastocyst stage and should not be performed earlier than 36 h after hCG treatment.     

In vitro-Fertilisationssystem der Ratte:Beeinflussung durch Ratten-Sper-matozoen-Antikorper — ein Experiment im allogenen System*

In vitro fertilization system of the rat. Influence of rat spermatozoal antibodies — an experiment in the allogenic system An in vitro fertilization model was set up using approximately three-months-old Wistar rats. Fertilization was carried out using Brinster medium with a pH of 7.8 and osmolarity of 280–320 m Osmol/l. Incubation took place at 37°C and with 5 % CO₂ in air. Ova were obtained by stimulating the female rats with 10 IU PMS and 30 IU HCG at intervals of 48 hours; after a further 15 hours the ova were then removed from the expanded oviduct. The inseminations were carried out with epididymal spermatozoa in pre-heated medium under paraffin-oil at a concentration of 1 × 10⁶ spermatozoa/ml. After 6 hours the ova were transferred into the culture medium corresponding to the fertilization medium. 48 hours after insemination the fertilization rate was ascertained on the basis of the two-cell stage. To the in vitro-fertilization cultures 10% (v/v) sera containing antisperm antibodies (immunization with rat sperm in Freund's Complete Adjuvant) were added. With this addition of antisperm antibodies a statistically significant reduction of the in vitro-fertilization rates in comparison to the control group (immunization with Freund's Complete Adjuvant) was achieved. Thus sera containing spermatozoa antibodies in vitro can be used to influence rats as well as mice, as has already been described for the latter. On the basis of this, an interesting model with regard to humans becomes conceivable, in which passive immunisations is carried out with spermatozoa antibodies, which are at present being produced in our laboratoy as monoclonal antibodies.