Seroprevalence of Toxoplasma gondii in sheep and goats in Rahim Yar Khan (Punjab), Pakistan.

Toxoplasmosis, an infection caused by Toxoplasma (T.) gondii Apicomplexa protozoan, is widespread in humans and other animal species, having already been reported in many countries and different climates. In Pakistan, no data is available on this aspect among food animals. This study was undertaken to determine the seroprevalence of T. gondii infection in sheep and goats. A total of 200 serum samples from sheep and goats, were collected from urban area of Rahim Yar Khan (Punjab), Pakistan and tested for Toxoplasmosis with a commercial latex agglutination kit (Eiken Chemical Co., Ltd. Japan). The overall seroprevalence of Toxoplasmosis was 19%. Goats had a significantly higher (p < 0.01) prevalence (25.4%) as compared to the sheep (11.2%); and higher (p < 0.01) in the female (24%) than in the males (19%) for both species. In the present study the male (both in sheep and goat) are found less seropositive T. gondii (OR = 0.23; 99% C.I. = 0.01, 1.81) as compared to female sheep and goat. The prevalence was significantly higher (p < 0.01) in adult sheep than younger animals. Among both the sheep and goats the group from 1-1.5 years are highly seropositive (OR = 1.75; 99% C.I. = 0.47, 6.51) as compared to the group less than one year of age followed by the 2-2.5 years age group (OR = 1.63; 99% C.I. = 0.50, 5.74) whereas group with more than 3 years of age least seropositive.

The effect of gamma interferon on IL-1 secretion of in vitro differentiated human macrophages.

After being cultured overnight, human monocytes lose their ability to secrete interleukin-1 (IL-1) when stimulated by lipopolysaccharide (LPS). However, when these monocytes were cultured for up to 9 days with various concentrations of interferon-gamma (IFN-gamma), these cells were found to retain their ability to secrete appreciable amounts of IL-1 on LPS stimulation. However, the effect was observed only if the monocytes were exposed to the IFN before LPS stimulation and simultaneous addition of IFN and LPS to macrophages was ineffective. This effect of IFN-gamma was related to the concentration of IFN added to the cultures and was completely neutralized by a monoclonal antibody to IFN-gamma. In addition to inducing IL-1 secretion, IFN-gamma also appeared to increase the overall production of IL-1, since reinduction of IL-1 secretion was not associated with a decrease in intracellular IL-1 content. When these macrophages were initially cultured with IFN-gamma, washed, and further cultured with IFN free medium, these macrophages were found to progressively lose their capacity to secrete IL-1 in response to LPS. Conversely, when monocytes were initially cultured in medium free of IFN, washed, and then further cultured in new medium, but now containing IFN-gamma, these macrophages were found to regain their capacity to secrete IL-1. However, the amount of reinduced IL-1 secretion decreased as the length of the initial culture period without IFN increased, with less than optimal IL-1 secretion occurring if monocytes were allowed to mature for 6 days before IFN-gamma pretreatment. In summary, these studies suggest that IFN-gamma may be important in enhancing IL-1 production and secretion by maturing macrophages and tissue macrophages and consequently may play a role in regulating the accessory cell activity of these cells for a variety of immune responses in vivo.

Failure of hydrocortisone to suppress the interferon-gamma-induced augmentation of interleukin 1 secretion of aged human monocytes.

The combined effect of hydrocortisone (HC) and interferon-gamma and -alpha (IFN-gamma and -alpha) on human blood monocytes (Mo) interleukin 1 (IL 1) secretion was investigated. IL 1 was generated by treating the fresh or aged Mo with lipopolysaccharide (LPS), and quantitated by radioimmunoassay (RIA). Hydrocortisone, at the pharmacological attainable concentration of 10(-5) molar (M), markedly suppressed fresh Mo IL 1 secretion but had no effect at lower tested doses. Addition of IFN-gamma enhanced the IL 1 secretion of fresh Mo; however, the simultaneous addition of 10(-5) M HC and IFN-gamma resulted in marked suppression of the monokine release. Monocytes, when cultured in vitro for three days, lost the capacity to secrete IL 1. The loss of IL 1 secretory potential of aged Mo was prevented by preincubating them with IFN-gamma prior to LPS stimulation. IFN-alpha was ineffective in this regard. Aged Mo, pretreated with the combination of IFN-gamma and HC were still able to secrete abundant quantities of IL 1, demonstrating the failure of HC to suppress the IFN-gamma-induced augmentation of IL 1 secretory potential. Even suprapharmacologic doses of HC (10(-4) M) did not inhibit this enhancement and actually further augmented it. Thus, therapeutic concentrations of HC suppress IL 1 secretion of fresh Mo even in the presence of IFN-gamma; however, therapeutic or suprapharmacologic concentrations of HC do not inhibit the IL 1 secretory capacity of IFN-gamma-treated aged Mo.

Role of IFN-gamma and alpha in IL 1 synthesis and secretion of in vitro differentiated human macrophages: a comparative study.

Human blood monocytes (Mo) cultured in vitro differentiate to macrophages (Mx) and lose the capacity to secrete interleukin 1 (IL 1) in response to endotoxin (LPS). Incubation of Mo with interferon gamma or alpha (IFN-gamma or IFN-alpha) prevented this loss of IL 1 secretory potential during the first 24 h of culture. However, there were marked differences between the two interferons if culture period was extended beyond 24 h. Incubation of Mo with IFN-gamma for 48 or 72 h induced IL 1 release in response to LPS in all the donors without exception. In contrast, 48-h incubation of Mo with IFN-alpha alpha caused IL 1 secretion (in response to LPS) in only a minority of donors, while 72-h incubation resulted in very little or no IL 1 release in all the individuals tested. Moreover, only IFN-gamma had the capacity to reinduce IL 1 secretory potential in Mx which had lost the capacity to secrete IL 1 during previous culture. These and other results suggest that IFN-alpha differs from IFN-gamma in being: a less potent IL 1 inducer, ineffective in maintaining IL 1 secretory capacity of fresh Mo for more than 48-72 h, completely unable to reinduce IL 1 secretory potential in culture-derived Mx. Thus, the two species of IFN appear to have a markedly different role in IL 1 synthesis and secretion.

Endobrst: simple and safe is best.

Minimal-access treatment for ureteropelvic junction (UPJ) obstruction is becoming increasingly complex. Is there still a place for the simple procedure of balloon dilatation? To examine the symptomatic and renographic results of patients at least 6 months after balloon disruption of an obstructed UPJ, we carried out a prospective audit of outcome as judged by DTPA renograms and clinical follow-up. The procedure has been performed on a total of 21 renal units (20 patients) with follow-up of 6 to 30 months (mean 22 months). Of these, 18 (86%) were asymptomatic at 3 months, although this success rate dropped to 17 (81%) by 6 months. Renographic split function improved in 11 renal units, and excretion improve in 14. Perioperative complications were few and associated with stent insertion. Three of the patients in whom the procedure failed went on to nephrectomy (14%); all had poor renal function (split <20%) at presentation. Although the quoted success rates are lower for endoluminal balloon rupture of stenosis treatment than open pyeloplasty, the former technique has significantly less morbidity. Our nephrectomy rate reflects our less than ideal early patient selection, and our balloon dilation technique is not recommended as a salvage procedure for patients in whom nephrectomy is inevitable. The learning curve is quick to climb, and this is a fundamentally simple procedure with medium-term results that remain encouraging.

The effect of IFN-gamma and alpha on Con-A stimulated T-cell proliferation: role of monocytes.

Interferon gamma (IFN-gamma) and alpha (IFN-alpha) have been shown to influence T-cell proliferation in several systems. Since both species of IFN affect monocyte (Mo) functions, it is conceivable that these effects are mediated through Mo. In the present study, the effect of IFN-gamma and alpha on concanavalin A (Con A)-driven T-cell proliferation was investigated under suboptimal culture conditions. T-cell proliferation was augmented by IFN-gamma. The stimulatory effect of IFN-gamma was Mo mediated and required the pretreatment of Mo with IFN-gamma prior to co-culture with T-cells. Addition of IFN-gamma during co-culture or pretreatment of T-cells with IFN-gamma was ineffective in achieving augmentation. IFN-gamma induced enhanced T-cell proliferation was not due to increased interleukin-I (IL-I) production by Mo pretreated with IFN-gamma, because addition of IL-I to control Mo-T-cells co-culture failed to increase T-cell proliferation. In contrast, IFN-alpha suppressed T-cell proliferation. The inhibitory effect of IFN-alpha was not mediated through Mo and could be achieved by preincubating the T-cells with IFN-alpha. These results demonstrate opposing effects of the two species of IFN on Con A-driven T-cell proliferation. In addition, Mo play an obligatory role in IFN-gamma induced stimulation, whereas Mo-IFN-alpha interaction is not required for IFN-alpha mediated suppression of T-cell proliferation.

Cottonseed with a high (+)- to (-)-gossypol enantiomer ratio favorable to broiler production.

This study was designed to evaluate the relative toxicity of (+)- and (-)-gossypol enantiomers in 0-3-week-old broilers. Treatments consisted of broiler starter diets formulated with either a glandless, which did not contain gossypol, a commercial glanded [62. 2% (+)-gossypol], or a glanded moco [83.2% (+)-gossypol] crushed cottonseed (CCS) (six replicates/treatment) plus a soybean meal negative control. Glandless cottonseed was mixed with the moco cottonseed (2.4% free gossypol) so that both the commercial glanded and moco glanded cottonseeds contained equivalent concentrations of free gossypol (2.0%). The cottonseed treatments were added at 5 and 10% of the diet. Body weights and feed conversions were determined weekly. Body weights and feed-to-gain ratios of broilers fed 5 and 10% glandless CCS and 5% moco CCS were not significantly different. Broilers receiving 10% commercial glanded CCS weighed significantly less than those subjected to all other treatments. Feed-to-gain ratios were significantly higher for broilers receiving 10% commercial glanded and 10% moco CCS as compared to 5% moco and glandless CCS, 10% glandless CCS, and control. Relative liver weights of birds receiving 10% moco CCS were significantly less than those of birds receiving 10% commercial CCS. The data clearly showed that broilers fed moco CCS containing a relatively high (+)- to (-)-gossypol enantiomer ratio performed better than broilers receiving commercial CCS with a lower (+)- to (-)-gossypol enantiomer ratio.

Time course evaluation of carotenoid and retinol concentrations in posthatch chick tissue.

Carotenoid and retinol concentrations were determined in chicks hatched from carotenoid-enriched eggs while being fed a carotenoid free starter diet. Single Comb White Leghorn (SCWL) hatching eggs were obtained from breeder hens fed diets enriched with either 0.2% beta-carotene, canthaxanthin, lutein, or basal control. Twenty-five day-old chicks hatched from each treatment group were placed in a battery and fed a carotenoid-free basal diet. At 1, 4, 7, 10, and 13 d, five chicks per treatment were randomly selected, and blood serum, liver, and unabsorbed yolk sac were collected for carotenoid and retinol analysis. There were no significant differences in retinol concentrations in the serum, liver, and unabsorbed yolk sac of the chickens. Only small amounts of beta-carotene were detected in the chick tissues. Concentration of all three carotenoids and retinol were greater in the liver and unabsorbed yolk sac than in serum. In the serum and liver, respective carotenoid concentrations decreased and retinol concentration increased with increasing age, whereas respective carotenoid and retinol concentrations in the unabsorbed yolk sac increased over time. It may be concluded that yolk sac carotenoids are not absorbed well, as unabsorbed yolk sac concentrations increased over time, and that carotenoid in serum and liver decrease linearly with increasing age.

Effect of beta-carotene, canthaxanthin, lutein, and vitamin E on neonatal immunity of chicks when supplemented in the broiler breeder diets.

The study was designed to assess neonatal immunity of chicks hatched from breeders fed diets supplemented with beta-carotene, canthaxanthin, lutein, or vitamin E. Broiler breeder birds were fed experimental diets consisting of control, 0.04% beta-carotene, 0.04% canthaxanthin, 0.04% lutein, 0.03% alpha-tocopherol acetate, or 0.04% beta-carotene plus 0.03% alpha-tocopherol acetate. Three weeks after initiation of experimental feeding, birds were vaccinated against Newcastle disease virus. Chicks hatched from the eggs of these breeders were used to determine the neonatal immune responses. There were no significant differences in weight gain and antibody titers of 3-wk-old chicks. 3H-Thymidine uptake by bursal lymphocytes when stimulated with tetrahydrofuran was significantly higher for the chicks hatched from breeders fed diets supplemented with vitamin E, or vitamin E plus beta-carotene, than in controls. 3H-Thymidine uptake by splenic lymphocytes when stimulated with concanavalin A and phorbol 12-myristate 13 acetate was significantly higher for the chicks hatched from breeders fed diets supplemented with vitamin E or beta-carotene alone, or vitamin E plus beta-carotene, than for the control chicks. Chicks hatched from hens supplemented with vitamin E had significantly higher antibody titers at 1 and 7 d of age than chicks from the control group. Vitamin E supplementation of breeder birds increased the immune response of their progeny.

Neonatal immune response and growth performance of chicks hatched from single comb White Leghorn breeders fed diets supplemented with beta-carotene, canthaxanthin, or lutein.

Forty Single Comb White Leghorn (SCWL) hens and 8 SCWL cocks were randomly divided into four treatment groups. Each group was fed a diet containing .02% beta-carotene, canthaxanthin, lutein, or basal control. After 20 d of feeding, eggs were collected daily from each experimental group for incubation. Two different hatches were set and chicks from each hatch were used for one of two different experiments. In both experiments, 24 chicks per treatment were vaccinated against Newcastle disease virus at 1 d of age and raised for 5 wk on a basal diet. In the second experiment, birds were revaccinated at 3 wk of age. In both experiments, at the end of 5 wk birds were killed and bursa of Fabricius, liver, and spleen were collected. For both experiments, there were no differences in antibody titers, weight gain, feed conversion ratio, and relative bursa weights of chicks. However in the second experiment, birds hatched from breeders fed lutein had significantly lower relative liver weights than chicks of the other treatments, whereas birds hatched from the breeders fed beta-carotene and canthaxanthin had significantly lower spleen weights than the control. These experiments suggest that carotenoids may not be effective in increasing neonatal immune response when they supplement practical breeder diets.

Tissue distribution of gossypol enantiomers in broilers fed various cottonseed meals.

Plasma and tissue concentrations of total, (+)- and (-)-gossypol were determined in broilers fed cottonseed meals (CSM) from eight oil mills (five expander solvent, two expeller, and one direct solvent). Free gossypol in the meals ranged from 0.033 to 0.180%, and total gossypol ranged from 0.974 to 1.459%. The (+)-enantiomer of gossypol varied from 53.8 to 61.3% of total gossypol. Eight CSM diets containing 28% CSM and a soybean meal control diet were fed to 162 1-d-old male broiler chicks during a 3-wk starter period. Concentrations of free gossypol in the CSM diets ranged from 92 to 504 microg/g, and total gossypol ranged from 2,626 to 4,085 microg/g. All diets were formulated with the same concentrations of digestible lysine and methionine and were fed ad libitum. At 21 d of age, there were no significant differences in body weights, feed conversions, or mortality of birds fed the CSM diets when compared to birds fed the soybean meal diet. Concentrations of (+)- and (-)-gossypol in plasma, liver, kidney, and muscle increased linearly as the level of free gossypol increased in the diets. Liver had the highest concentration of total gossypol (71.4 to 313.6 microg/g DM) followed by kidney (9.2 to 36.3 microg/g DM), plasma (3.0 to 14.6 microg/mL), and muscle (2.1 to 9.8 microg/g DM). The proportion of (-)-gossypol was higher in plasma (26.7%) and kidney (25.6%) than in muscle (19.1%) and liver (16.0%). Performance data from this study indicate CSM can be used successfully in chick starter diets at levels up to 28% when diets are formulated on a digestible amino acid basis.

Use of expander cottonseed meal in broiler diets formulated on a digestible amino acid basis.

This experiment was designed to evaluate the use of expander cottonseed meal (CSM) in broiler diets formulated on a digestible amino acid basis and to determine the tissue distribution of gossypol enantiomers and their relationship to gossypol consumption. Gossypol is an antinutritional polyphenolic pigment found in cottonseed. A total of 800 1-d-old broilers (Arbor Acres) was randomly divided into 40 groups of 20 birds each and placed in a broiler house with wood shavings litter. The CSM was included in starter and grower diets at 0, 7, 14, 21, and 28% of the diet. Diets were formulated to have similar levels of digestible methionine and lysine as the 0% CSM control diet. At 21 d of age, five birds per pen were randomly selected for the determination of gossypol enantiomers in plasma, liver, heart, and breast muscle. Tissues were again collected at 42 d of age. Results of this experiment indicated that at 21 d of age cumulative body weight and feed-to-BW ratios of birds fed CSM diets were not significantly different from the control. By 35 d of age, feed-to-BW ratios of experimental birds fed the 28% CSM diet were significantly greater than the control birds, and by 42 d, BW were lower than those of the control group. Tissue concentrations of total, (+) and (-) gossypol increased linearly as the level of CSM increased. At 42 d, liver had the most gossypol with a ratio of 87% (+) gossypol to 13% (-) gossypol. Plasma contained 73% (+) gossypol and 27% (-) gossypol. Heart contained 45% (+) gossypol and 55% (-) gossypol. Breast muscle had the least gossypol. Results of this experiment indicated that expander solvent CSM could be fed to broilers at up to 21% of the total diet if amino acid digestibility was considered.

Evaluation of sorghum ergot toxicity in broilers.

Three experiments evaluated the performance of broilers fed sorghum ergot consisting of sphacelia/sclerotia of Claviceps africana present in tailings removed by conditioning of seed from grain sorghum hybrid seed production fields near Uvalde (Experiments 1 and 2) and Dumas (Experiment 3), Texas. Percentage sphacelia/sclerotia and total alkaloid content, respectively, in sorghum ergot tailings were 8% and 11.3 ppm for Uvalde and 75% and 235 ppm for Dumas. Sorghum ergot and control sorghum diets were based on the NRC (1994) requirements for starting broilers. In Experiment 1, neither growth nor feed efficiency were significantly reduced in male broilers fed sorghum ergot from hatch to 3 wk of age, but liver weights were significantly greater than those in the control. In Experiment 2, straight-run broilers were raised to 6 wk of age in floor pens using a three-phase feeding program. Sorghum ergot significantly reduced gain in 4-wk-old broilers and cumulative body weight at 5 wk. Feed conversion was significantly reduced during all three phases of feeding. In Experiment 3, control sorghum and the 75% ergot tailings were added to corn-soy basal diets at rates of 2.5, 5, and 10% by weight and fed to male broilers from hatch to 3 wk of age. Sorghum ergot did not significantly reduce growth, but, during Weeks 2 and 3, feed-to-gain ratios were higher. Neither type nor concentration of sorghum ergot significantly affected relative liver weights. We did not observe significant mortality or obvious symptoms of ergot toxicity, such as necrotic lesions of the feet or vesicular dermatitis of the comb, in any of the three experiments.

Prion protein gene polymorphisms in four goat breeds of Pakistan.

Four different goat breeds (Pak-Angora, Dera Din Panah, Naachi and Teddy) of Pakistan were selected to investigate polymorphism in the prion protein gene (PrP gene) responsible for scrapie disease resistance in goats. Initially, genotyping of 187 animals of these four breeds by restriction fragment length polymorphism (RFLP) was done to see the genotype for codon 136 and 154. All the animals were monomorphic with a genotype of AARR except one animal of Teddy breed having the genotype of AARH. Sequencing of PrP gene of twenty animals representing these four goat breeds revealed two genotypes PPSSSS and PPSSPS with haplotypes PSS and PSP of PrP gene at the codon numbers 42, 138, and 240. All four breeds showed both wild type monomorphic sequence and mutant polymorphic sequences of these codons. The mutants of 42 and 138 codons translate the same amino acids as with the wild type sequences, while the mutant of codon 240 is responsible for a different amino acid translation i.e., serine to proline. In short, this study provides preliminary information about alleles and genotypes of PrP gene in four goat breeds of Pakistan.

Field evaluation of Eimeria tenella (local isolates) gametocytes vaccine and its comparative efficacy with imported live vaccine, LivaCox.

The present paper describes the field evaluation of local gametocyte vaccine and its comparative efficacy with commercial anticoccidial vaccine, LivaCox, used in breeder and broiler flocks in Pakistan. Humoral immune response in vaccinated and control chickens was monitored by enzyme-linked immunosorbent assay. Results demonstrated significantly elevated antibody titres in vaccinated groups as compared to control groups conducted both in Laboratory and field experiments. Significantly (P < 0.01) higher antibody titres in local gametocyte-vaccinated group as compared to LivaCox-vaccinated chickens were recorded. Splenic cell migration inhibition assay was used to detect the cell-mediated immune (CMI) response, and results were expressed in terms of per cent migration index. Lower per cent migration index in LivaCox-vaccinated chickens indicated the higher CMI response, as compared to local gametocyte-vaccinated chickens, although the difference was statistically non-significant (P > 0.05). Results of the challenge studies in laboratory experiments revealed significantly higher (P < 0.05) oocyst count in LivaCox-vaccinated group as compared to local gametocyte-vaccinated chickens.Maximum protection (75%) against mixed species of genus Eimeria was recorded in chickens vaccinated with gametocyte vaccines as compared to LivaCox-vaccinated group. The mean body weight gains in chickens vaccinated with local gametocyte vaccine were significantly better (P < 0.05) than in chickens vaccinated with LivaCox vaccine, both in laboratory and field experiments. Majority of the chickens (70-72%) in control group demonstrated severe lesions (3.0-4.0), while 20-26% chickens showed moderate lesions (2.0). On the other hand, local gametocyte- and LivaCox-immunized chickens developed 78% and 85% mild to moderated lesions (1.0-2.0), respectively. Results of the present study provide a probable explanation for cross-protection induced by Eimeria tenella gametocyte vaccines against other species of genus Eimeria.

Transformation of polycythemia vera to Ph-positive chronic myelogenous leukemia.

A patient with polycythemia vera (PV) developed chronic myelogenous leukemia (CML) 8 years after his initial presentation with erythrocytosis. The patient received a total of 3.5-7 g of chlorambucil over an approximate period of 5 years. Transformation of PV to CML is very unusual; only three substantiated cases had been reported in literature until 1975, and none has been described since then. All patients received either 32P or chemotherapy for PV treatment, suggesting that irradiation and alkylating agents may have played a role in the transformation to CML.

1,25-Dihydroxyvitamin D3 (calcitriol) suppresses IL-2 induced murine thymocyte proliferation.

1,25-dihydroxyvitamin D3 (Calcitriol) inhibits mitogen stimulated T cell proliferation by blocking the production of Interleukin-2 (IL-2). The present study was initiated to determine whether the action of calcitriol was limited only to inhibition of IL-2 production, or if it influenced other events as well. To avoid the use of lectins, thymocytes from CD-I Swiss mice were chosen, which proliferate in response to Interleukin-I (IL-I) or IL-2 without the addition of lectins. Calcitriol inhibited (80-90%) IL-I induced CD-I mouse thymocyte proliferation, whereas 25-OH-D3 was ineffective. Further addition of IL-I failed to overcome this suppression. Surprisingly, calcitriol also inhibited IL-2 induced thymocyte proliferation (60-80%). Further addition of IL-2, in the presence of calcitriol, was ineffective in enhancing thymocyte proliferation. Similar results were obtained with C57BL/10 mouse thymocytes. Additional studies excluded the possibilities that calcitriol mediated inhibition was due to calcitriol IL-2 binding or damage to thymocytes by calcitriol. Thus, calcitriol not only blocks IL-2 production, but these results strongly suggest, that it also interferes with IL-2-thymocyte interaction, which in turn results in inhibition of thymocyte proliferation.