Histopathology and Immunohistochemistry Assessments of Acute Experimental Infection by <i>Brucella melitensis</i>in Bucks

Background: Brucellosis in male goats is characterized by arthritis, orchitis and epididymitis, which may induce infertility. Nevertheless, these lesions were categorized as chronic while acute lesions had not been described. This study investigates the histopathological and immuno histochemistry reactions in organs of bucks acutely infected by Brucella melitensis. Results: Only testis and prepuce of acutely infected bucks showed significantly severe histological lesions. Other internal organs had mild to moderate lesions. However, positive immunohistochemistry stainings were observed in organs except the bulbourethral gland. There was a significant positive correlation between the distribution of B. melitensis and IHC intensity but no significant correlation between the IHC intensity and histopathology lesions. Conclusion: The results indicate that acute brucellosis did not lead to clinical presentation, although B. melitensis was well distributed in various organs of bucks.

Genital Tract Morphopathology of Red Sokoto and West African Dwarf Does in Makurdi

This study was conducted with the aim to record pathological conditions of the female genital tract of Red Sokoto (RS) and West African Dwarf (WAD) does with the objective of comparing the prevalence of genital tract pathology between the two breeds. Of the 211 (RS) and 221 (WAD) genital tract collected and examined, 104 (49.3%) of RS and 97 (43.3%) of WAD showed different types of pathological conditions. Highest occurrence of pathological conditions found in RS was in the uterus (38.4%), followed by those in the ovary (30.8%), cervix (6.6%), salpinx (6.2%) and vagina (1.4%) while in WAD, pathological conditions were highest in the uterus (28.1%), and followed by ovary (26.8%), salpinx (5.0%), cervix (2.2%) and vagina (0.5%). In RS and WAD does, respectively, endometritis (11.9% and 7.1%) was the most common pathology followed by ovaro-bursal adhesion (10.4% and 5.4%), inactive ovaries (4.3% and 4.9%), cystic Graffian follicles (4.2% and 2.2%), paraovarian cyst (3.3% and 3.1%), uterine haemorrhage (3.8% and 2.7%), and ovarian hypoplasia (2.4% and 4.5%). Other pathological conditions found in RS and WAD does include cervical lesions: cervicitis (2.8% and 1.8%), haemorrhage (3.2% and 0.5%);uterine lesions: hydrometra (1.9% and 1.3%), pyometra (2.8% and 1.8%), mucometra (2.8% and 3.6%), metritis (2.4% and 1.3%), post parturient metritis (2.8% and 1.8%), caruncular atrophy (1.4% and 0.5%), endometrial hyperplasia (1.0% and 1.8%), melanosis (1.9% and 1.8%), post parturient emphysematous metritis (1.4% and 2.2%), perimetritis (1.4% and 2.2%), uterine congestion (1.9% and 2.2%), oedema (1.0% and 0.5%) and Cysticercus tenuicolis cyst (0.5% and 0.5%);esophageal lesions: salpingitis (1.9% and 3.1%), hydrosalpinx (1.0% and 0.5%), tubo-bursal cyst (3.3% and 0.9%), ovarian lesions: cystic corpora lutea (2.8% and 1.8%), oophoritis (0.5% and 1.8%), and Cysticercus tenuicolis cyst (1.0% and 0.9%). Hyperaemia (0.5%) and calcification of cervical caudal rugae (0.5%) occurred in RS only. Genital lesions were not statistically different (P > 0.05) between RS and WAD does. In the two breeds, the 13 - 24 months age group had high prevalence of genital lesions. The current study has shown that endometritis and ovaro-bursal adhesions are the common female genital tract pathological conditions in Red Sokoto and West African Dwarf does. The high prevalence of female genital tract abnormalities may suggest that genital diseases are of serious limitation to goat production in Makurdi.

Physio-pathology of induced endotoxaemia in bovine and its treatment regimen

Endotoxic shock was induced in five apparently healthy male buffalo calves by i.v infusion of Escherichia coli endotoxin at 5microgram/kilogram (μg/Kg) body weight/hour (BW/hr) for 3 hours. Endotoxin infusion caused clinical signs of restlessness, respiratory distress, snoring, diarrhoea, profuse salivation along with the significant hypoproteinemia, hypoalbuminemia and hypokalemia in all the animals. The animals were observed up to day 4 or death, whichever was earlier. The treatment with one time intravenous infusion of hypertonic saline solution @ 4milliliter/Kilogram body weight (ml/Kg?BW), dextran-40 @ 10 ml/Kg?BW, flunixin meglumine @ 1.1 milligram/Kg?BW (mg/Kg?BW) and blood @ 20 ml/Kg?BW to these animals alleviated the clinical signs and significantly raised the circulating glucose level at 4.5 and 5.5 hrs. The treatment led to survival of three of the five endotoxemic buffalo calves. The significant hypoproteinemia, hypoalbuminemia, hypokalemia and hypoglobulinemia continued even after treatment. Gross and histopathologic findings of congestion, haemorrhage, necrosis in vital organs viz., lungs, liver, kidneys, brain and intestines were suggestive of endotoxin induced hypoxia and multi-organ failure. Additionally, emphysema and fibrinous thrombi in microvasculature of lungs were salient histopathological findings indicating terminal respiratory failure in the remaining two dead endotoxemic buffalo calves. From clinical signs, plasma chemistry and pathological lesions, it was concluded that endotoxemia led to a disruption of critical life processes, but a timely and effective treatment could counter these deleterious effects and save precious lives.

Alleviating effects of melatonin on oxidative changes in the testes and pituitary glands evoked by subacute chlorpyrifos administration in Wistar rats

Objective:To evaluate the alleviating effects of melatonin on oxidative changes in the testes and pituitary gland induced by subacute chlopyrifos（CPF） exposure in rats.Methods:Forty adult male Wistar rats divided into 4 groups of 10 animals were used for the study.Croup I received soya oil（2 mL/kg） while group II was administered with melatonin（0.5 mg/kg）.Group III was administered CPF only(8.5 mg/kg l/10th of the LD50) while group Ⅳ was pretreated with melatonin（0.5 mg/kg） and then exposed to CPF（8.5 mg/kg）,10 min later.The regimens were administered by gavage once daily for a period of 28 d.At the end of the exposure period, the rats were sacrificed and the testicular tissues and pituitary glands were evaluated for the malonaldehyde（MDA） concentration and activities of superoxide dismutase（SOD） and catalase（CAT）.Results:CPF increased MDA concentrations and reduced the activities of SOD and CAT in the testes and pituitary gland.Melatonin pretreatment reduced the testicular and pituitary MDA concentrations and improves the SOD and CAT activities.Conclusions:the study showed that subacute CPF-induced oxidative stress in the testes and pituitary glands were alleviated by melatonin due to its antioxidant property.

Induction of Apoptosis in the Immature Mouse Testes by a Mixture of Melamine and Cyanuric Acid

The toxicity of melamine has attracted much attention since the recent outbreaks of renal injury in pets and infants. Previous studies indicated that melamine by itself had low toxicity, whereas a mixture of melamine and cyanuric acid （M＋CA） could cause serious renal damage. At present, most researches on the toxicity of M＋CA are focused on the kidney. However, little is known about the adverse effects of this mixture on the reproductive system. In the present study, the toxicity of M＋CA to testes was investigated. Immature male mice were orally dosed with 0, 0.6, 3, and 15 mg kg-1 d-1 of a 1：1 M＋CA for 28 d. Pathological changes occurred in germ cells, such as loose arrangement, reduced numbers and karyopyknosis, indicating that this mixture was toxic to spermatogenesis. Compared with the control group, the TUNEL- positive germ cells increased significantly and the ratio of Bcl-2 to Bax, total antioxidant capacity and superoxide dismutase activity decreased significantly in the 3 and 15 mg kg-1 d- M＋CA treated group, while the activities of caspase-3, caspase- 8 and caspase-9 remained unchanged. The results suggest that M＋CA can induce apoptosis in the mice testes. The downregulation of Bcl-2/Bax and oxidative stress may play a pivotal role in the induction of apoptosis by M＋CA in mice testes.

Aldehyde dehydrogenase activity helps identify a subpopulation of murine adipose-derived stem cells with enhanced adipogenic and osteogenic differentiation potential

AIMTo identify and characterize functionally distinct subpopulation of adipose-derived stem cells (ADSCs). METHODSADSCs cultured from mouse subcutaneous adipose tissue were sorted fluorescence-activated cell sorter based on aldehyde dehydrogenase (ALDH) activity, a widely used stem cell marker. Differentiation potentials were analyzed by utilizing immunocytofluorescece and its quantitative analysis. RESULTSApproximately 15% of bulk ADSCs showed high ALDH activity in flow cytometric analysis. Although significant difference was not seen in proliferation capacity, the adipogenic and osteogenic differentiation capacity was higher in ALDHHi subpopulations than in ALDHLo. Gene set enrichment analysis revealed that ribosome-related gene sets were enriched in the ALDHHi subpopulation. CONCLUSIONHigh ALDH activity is a useful marker for identifying functionally different subpopulations in murine ADSCs. Additionally, we suggested the importance of ribosome for differentiation of ADSCs by gene set enrichment analysis.

Food Hygiene Knowledge, Attitudes and Practices of the Food Handlers in the Military Hospitals

Improper practices and lack of knowledge by food handlers are contributing factors for the spread of foodborne outbreaks. This study aimed to explore the knowledge, attitude and practice of food handlers in military hospitals in Jordan. A Self completed questionnaire was answered by 200 employees;150 military employees and 50 civilian employees in 7 Jordanian military hospitals selected randomly one from the capital Amman and two from the three provinces. The results showed that the means of the percentage scores for the knowledge, attitude, practice, were 84.82, 88.88, 89.43, respectively and the overall knowledge, attitude, practice (KAP) mean percentage score was 87.88. A significant difference (P 0.05) in these values was observed between the military and civilian employee. The mean percentage scores for knowledge, attitude and practice were 88.5, 93.2, and 91.4, respectively for military employee compared to those for civilian employee of 73.7, 75.8, and 83.5, respectively. A significant difference for the overall (KAP) mean percentage score was also observed between gender, education and specific occupation of the food handler. Female KAP percentage mean score was 90.0 and that for males was 86.6;The college or university educations employee had a mean score of 91.6 while the elementary school education employee had a mean score of 79.7;The nutritionists and cooks had statistically similar means of 91.6 but these occupations significantly differ from the waiters 84.4 and cleaners 80.2.

Diazinon-Induced Clastogenity and Pathological Changes in Ovaries and Testes of Clarias gariepinus

In most tropical developing countries, one of the problems facing aquaculture industry is the pollution of ponds and rivers with pesticides. Chemicals such as diazinon, an organophosphate pesticide, originating from agricultural activity enter the aquatic environment through atmospheric deposition, surface run-off or leaching. Pollutants enter the food chain through accumulation in soft bottom sediment and aquatic organisms. However, information on how these pesticides affect inhabiting organisms is often not available. In a triplicate experimental set-up, seventy-two (72) apparently healthy catfish comprising adult and juvenile of both sexes were therefore exposed to a previously determined no effect concentration (0.405 ppm) of diazinon. Another set of fish was exposed to 0.0625 μg sodium arsenite, a known clastogen, which was used as the positive control, while another set of catfish exposed to the culture water alone was the negative control. Adults and juveniles were exposed separately to avoid cannibalism. After 48 hours of exposure, micronuclei induction was determined in subsets of experimental groups, while exposure continued for 28 days. Catfish organs were harvested on days 21 and 28 to determine the effect of long-term exposure to diazinon on histology. Water quality was also monitored before and during exposure in the experimental groups. The result established a significantly high mean micronucleated polychromatic erythrocytes (15.00) in catfish exposed to diazinon suggesting genetic damage (normal is ≤4). The MPE in sodium arsenite exposed fish was 28, while that of the control group was below 4. Effect of sex and age on micronuclei induction was not significant. Histological alteration observed in the ovary and testis was distorted matured cells and extensive testicular degeneration, respectively. The results show that diazinon has clastogenic effect, and may have endocrine disrupting properties because of the histological changes induced in the ovaries and testis.

Bacteriological and pathological studies of egg peritonitis in Commercial layer chicken in Namakkal area

Objective:To detect the various bacteriological agents and pathological changes in commercial layer chicken affected with egg yolk peritonitis in Namakkal region of India.Methods:A total of 6572 layer chicken from 85 commercial farms were subjected for the study,out of which 1715 showed various types of oviduct almoimalities.Among the 1715,264 birds from six farms were identified as egg peritonitis on the basis of postmortem examination.Trachea,lung,heart blood,liver,peritoneal exudate,oviduct(infundibulum,magnum,uterus)and cloacal swabs were collected from the 264 birds with egg peritonitis lesion for screening of bacterial agents.Signalment,clinical signs and pathological changes were recorded in the affected flocks.Result:The results of the present investigation indicated that the E.coli associated egg peritonitis was responsible for 15.39%of the reproductive tract abnormalities in commercial layers between 21 and 80 week of age.In the affected flocks egg production drop and mortality varied from 3%to 20%and 0.5%to 7.0%respectively.It was noticed during peak egg production(21 to 60week)and southwest monsoon season(58%).Statistical analysis of age,season and egg production by Chi square test of independence revealed highly significant difference.E.coli was isolated as a pure culture and concurrent with other bacterial agents in 226 and 38 birds respectively.Among the fifteen E.coli serotypes identified serotype O_(166),O_(?)and O_(111)were predominant.Necropsy examination of affected birds revealed the presence of amorphous or insipissiated yolk material in the abdominal cavity with inflammatory changes in the ovary,oviduct and intestine.Microscopically the oviduct surface epithelium showed degeneration and desquamation,moderate to marked infiltration of inflammatory cells especially heterophils and lymphocytes in various regions and lumen contained serofibrinous exudate,inflammatory and desquamated epithelial cells with bacterial microcolonies.Ovarian follicles revealed hyperemia,degeneration of granulosa cells and infiltration of inflammatory cells.Intestine showed degenerative,necrotic and inflammatory lesion.Conclusion:The findings of this study showed that the egg peritonitis might be caused by either the translocation of intestinal E.coli into the peritoneal cavity or by the movement of cloacal E.coli into the oviduct followed by ascension of these bacteria up the oviduct,through the infundibulum,and into the peritoneal cavity.To control the egg peritonitis faecal contamination with E.coli should be minimized.

Nationwide seroprevalence, spatial distribution and risk factors of Leishmania in Jordan

Objective:To explore the seroprevalence,spatial distribution and risk factors for Leishmania seropositivity in Jordan.Methods:Blood samples from 872 apparently healthy participants were randomly selected from 11 governorates in Jordan and tested for anti-Leishmania K39 IgG.Risk factors(animal ownership and agriculture practices)and demographic data were also collected using pre-tested and validated questionnaire.Results:Overall,2.52% of participants were seropositive for Leishmania spp.Participants living in the Jordan Valley plateau had significantly greater odds(adjusted odds ratio= 3.70,95% CI 1.37-9.93)of seropositivity than those living in the Highlands after adjustment for age.Conclusions:This study supports the intermittent reports of cutaneous leishmaniasis outbreaks in the Jordan Valley.Vector control measures in the Jordan Valley should be considered,including insecticide treated bed nets,sugar baits and using flowering plants to attract and trap Phlebotomus papatasi sand flies.Active surveillance in the Jordan Valley is also recommended in light of this and other reports.

Pathogenesis of Demyelinating Encephalopathy in Dogs with Spontaneous Acute Canine Distemper

So far, the pathogenesis of demyelination caused by canine distemper virus （CDV） in the central nervous system has remained unclear, although a lot of studies have been done extensively. To further investigate the relation of variety cells in brain to demyelination, this study was performed on 15 dogs with spontaneous acute canine distemper and 2 controls. According to anatomical relation, the brain was divided into cerebrum, cerebral stem and cerebellum. The sections with no, mild, moderate, or severe demyelinating lesions were selected respectively and stained by HE and immunohistochemistry. Immuno-localisation of CDV antigen was used to conftrm CDV infection. The brain was examined for co-localisation of the CDV antigen with either an astrocyte-specific marker, glial fibrillary acidic protein （GFAP）, or an oligodendrocyte-specific marker, galactocerebroside （GalC）. Apoptotic cell was detected by TdT-mediated nick end-labeling assay （TUNEL）. The results demonstrated that the local disturbance of blood circulation mainly included congestion, edema, thrombosis, and disseminated intravascular coagulation （DIC）. The CDV neucleocapsid protein positive reaction, metabolic disorder and apoptosis of oligodendrocytes were observed in demyelinating areas. Lots of astrocytes displayed CDV antigen-positive, especially in their process. Some of them became apoptotic cell confirmed by TUNEL staining. Fibrous astrocytes showed more intense GFAP-positive in mild and moderate demyelinating area. Some of nervous cells located in pyramidal cell layers and nucleus nervi were in degeneration, necrosis. Satellitosis, neuronophagia and apoptotic neurons were examined by hematoxylin and eosin （HE） and TUNEL staining. The results suggested that the demyelinating changes in brain tissues infected with CDV mainly related to the metabolic disorder and apoptosis of ogliodendrocytes and astrocytes; also involved with the local disturbance of blood circulation and some neuron lost.

Experimental Trypanosoma brucei infection at immediate post partum period:effects on dam and the offspring

Objective:To investigate the effects of immediate post-partum infection with Trypanosoma brucei（T.brucei） on dam and offspring.Methods:Sixty female Albino rats（Rattus norvegicus） weighing between 130-170 g were used as animal model.The animals were divided as follows: 25 infected between 1-5 days post partum;10 infected unbred as positive controls;and 25 uninfected as negative controls.The following parameters were evaluated:packed cell volume （PCV）,level of parasitaemia,survival time,litter size and litter weight at birth and on days 7, 14 and 21 post delivery,using conventional methods.Possible trans-mammary transmission of infection to litter through milk was also assessed.Results:The results showed a comparatively （P【0.05） higher mean PCV value for the uninfected negative control on the 8 day post infection compared with the infected groups which corresponded with the increasing level of parasitaemia in the two infected groups.Mean litter size and litter weights were higher（P【0.05） in the uninfected controls on the 21st day.Survival time in the infected groups were similar.No evidence of trans-mammary transfer of infection was recorded.Conclusion:T.brucei infection during immediate post partum period is detrimental to the dam and impairs growth of the offspring.

Methanolic extract of Clausena excavata promotes wound healing via antiinflammatory and anti-apoptotic activities

Objective: To investigate the anti-inflammatory properties of methanolic extract of Clausena excavata in lipopolysaccharide(LPS)-activated macrophages(J774 A.1) and the effect on skin wound in a rat model through determining cytokine levels and gene expressions. Methods: The effects of methanolic extract of Clausena excavata on in vitro viability and TNF-α, IL-6, IL-10, and nitric oxide release by LPS-activated J774 A.1 cells were determined. In addition, relative expressions of BAX, BCL-2 and COX-2 genes were examined in healed wounds of rats. Results: The methanolic extract of Clausena excavata was not toxic to J774 A.1 cells at the highest dose of 400 μg/m L. It decreased levels of TNF-α and IL-6, while increasing IL-10 level in LPSactivated J774 A.1 cells and in the healed wounds of rats. The methanolic extract of Clausena excavata also inhibited nitric oxide production in LPS-activated J774 A.1 cells. The BAX and COX-2 genes were downregulated while the BCL-2 gene was upregulated in the healed wound of rats. Conclusions: The methanolic extract of Clausena excavata promotes wound healing via its anti-inflammatory and anti-apoptotic activities.

Evaluating Feed Intake of Fishes in Aquaculture Nutrition Experiments with Due Consideration of Mortality and Fish Survival

Mortality is normal and natural in aquaculture experiments. However, it becomes a problem when measuring feed intake as a component of nutrient utilization parameters such as： feed conversion ratio, feed conversion efficiency, apparent net protein utilization, protein efficiency ratio and protein productive value. In order to accurately evaluate feed intake with due consideration of mortality encountered, a formulae was developed using empirical data obtained from an experiement that had mortality. The formulae Fwas： total feed consumed by numbers of survived fishes=DL∑i=1n（Fi/Ni）N. Where, D=Numbers of days fishes were fed within each period ibefore it was adjusted（e.g fortnight）, it was constant throught period of experiment; F=Uantity of feed fed per day for a fortnight before the quantity was adjusted, it changed every fortnight; L=Numbers of living fishes at the end of experimental period; N=Numbers of fishes at each weighing period, it might change every fortnight or might be constant if no mortality occurred; while n=Numbers of times fishes were weighed and quantity of feed was adjusted but this did not include the final weighing at the termination of experiment. However, if the fishes were weighed weekly, then D=6.

A Combination of Antigen Retrieval Methods Significantly Enhances the Unmasking of Tedious Antigens of Lymphangiogenesis and Angiogenesis in Canine Mammary Tumour

The use of immunohistochemical techniques has enhanced the diagnostic intensities in pathology. The sensitivity and specificity of any immunohistochemical protocol mainly depends on the ability to unmask antigen from formalin fixed tissue. Various combinations of antigen retrieval protocols have been used to unmask antigen efficiently but none has been established as a flawless one. These protocols either have one or the other lacunae, lacking repeatability under same conditions. In our study, we tried to develop a standardized immunohistochemical protocol for the identification of blood and lymphatic vessels in tissue sections of canine mammary tumour (CMT) using recently identified markers. The combined effects of antigen retrieval (AR) methods including pH shock by Heat-induced epitope retrieval (HIER) and Protease-induced epitope retrieval (PIER) were found highly effective in retrieval of tedious antigens of lymphangiogenesis and angiogenesis. The use of combined antigen retrieval technique for the unmasking of antigens from over fixed, old formalin fixed paraffin embedded (FFPE) canine mammary tumour sections resulted in efficient unmasking of epitopes when compared with the individual antigen retrieval methods.

Effect of Mineral and Vitamin Supplementation on Performance and Haemotological Values in Broilers

A total of 120-day-old broilers were divided into four groups. In group-A, broilers were left without supplementation of minerals and vitamins(control), while the broilers in groups B, C and D were given mineral and vitamin supplementation 1-6, 1-4 and 5-6 weeks, respectively. The results showed that the feed consumption of broilers in groups A, B, C and D was 4 234.61, 4 028.38, 4 169.32 and 4 181.87 g · b-1; water intake was 9 668.88, 9 295.55, 9 882.11 and 9 402.86 L · b-1; live body weight was 2 068.90, 2 360.90, 2 255.30 and 2 157.40 g · b-1, respectively. The feed conversion ratio of broilers in groups A, B, C, D was 2.05, 1.71, 1.85 and 1.94 g · b-1, carcass weight(wt) was 1 550, 1 656, 1 552 and 1 528.8 g · b-1, and average dressing percentage was 61.24, 63.02, 62.60 and 60.51 respectively. Weight of heart in groups A, B, C and D was 15.13, 13.16, 11.72 and 11.50 g · b-1, weight of gizzard was 32.98, 37.48, 37.62 and 33.35 g · b-1, weight of liver was 51.94, 48.70, 51.94 and 48.70 g · b-1 and spleen weight was 3.48, 3.28, 3.06 and 2.82 g · b-1, respectively. Average mortality of broilers in groups A, B, C and D was 6, 2, 4 and 5 resulting 20.00%, 8.00%, 13.00% and 16.00% mortality rate, respectively. The haemoglobin level in blood of broilers in groups A, B, C and D was 9.20, 12.23, 10.87 and 7.60 g · dL-1,white blood cell level 77.47, 89.83, 81.63 and 78.13(×106 μL); red blood cell level 1.85, 2.58, 2.45 and 1.40(×106 μL), respectively. It was concluded that broilers given mineral and vitamin supplementation(Aquavit Electrolyte) for 6 weeks were proven to be most effective regime to result the lowest feed intake, the highest live body weight, better feed conversion ratio, higher carcass weight, better dressing percentage, higher net profit, relatively improved haemoglobin level in blood, increased counts of red blood cells and white blood cells. In all the above parameters, the broilers received mineral and vitamin supplementation for the first 4 weeks and the last 2 weeks showed relatively inferior performance, it is suggested that mineral and vitamin supplementation through water for partial period was not much effective in broiler productivity. Broilers may be supplemented with commercially available minerals and vitamins in drinking water over full rearing period, for achieving higher live body weight and efficient feed conversion ratio.

A novel mouse model of adult T-cell leukemia cell invasion into the spinal cord

Adult T-cell leukemia( ATL) is a mature T-cell malignancy caused by human T-cell leukemia virus type I infection, and 10%-25% of patients show central nervous system( CNS) involvement. CNS involvement significantly reduces survival and there are no effective treatments for CNS involvement. Therefore, an appropriate animal model is required to evaluate the inhibitory effects of novel drugs on the progression of ATL with CNS involvement. Here, we established a mouse model of ATL with CNS involvement using NOD.Cg-Prkdc~ (scid) Il2 rg ^(tm1Wjl)/SzJ mice inoculated with ATL cells intramuscularly in the postauricular region, and these mice showed paraparesis. Of the 10 mice inoculated with ATL cells intramuscularly(I.M.) at 5 weeks of age, 8(80%) showed paraparesis, whereas none of the 10 mice inoculated with ATL cells subcutaneously(S.C.) showed paraparesis. In the I.M. group, PCR detected HTLV-1-specific genes in the thoracic and lumbar vertebrae; however, in the S.C. group, the vertebrae were negative for HTLV-1 genes. Histological analysis revealed a particularly high incidence of tumors, characterized by accumulation of the injected cells, in the thoracic vertebrae of mice in the I.M. group. Tumor cell infiltration was relatively high in the bone marrow. Spinal cord compression caused by invasion of the tumor mass outside the pia mater was observed in the thoracic vertebrae of the spinal cord. In conclusion, we have reported a mouse model of tumor growth with paraparesis that may be used to assess novel therapeutic agents for ATL with CNS involvement.

Diagnosis of Clinical Cases of Infectious Bursal Disease Using a Modified Rapid Taq Man-MGB Real-Time RT-PCR Assay

Infectious bursal disease(IBD)is an important contagious viral infection of immune system of poultry.This infection possesses a permanent threat to the profitability of poultry industry worldwide.The aim of this work was to modify the Taq Man-MGB real-time reverse transcription-polymerase chain reaction(rRT-PCR)in one step involving two fluorogenic Taq Man labeled probe and using this protocol for detection of infectious bursal disease virus(IBDV)collected from suspected cases distributed in different regions of the country during the period 2013-2016.The intralaboratory validation of modified method was realized for specificity,linearity,repeatability,sensitivity and reproducibility.It allowed reducing the test running time by six folds.This method was applied on 102 pools of bursa of fabricius(BF)samples collected from affected broiler farms suspected to be infected by IBDV.Birds showing macroscopic lesions including muscle petechial hemorrhages,hypertrophy and hemorrhage of BF,were subjected to molecular analysis using modified protocol“Taq Man-MGB rRT-PCR”.The validation satisfied all criteria and the assay developed could be a useful tool for a very rapid diagnosis of IBDV and permit to detect and to discriminate in one-step very virulent(vv)from non-vv(classic and variant)IBDV strains.Out of 84 IBDV positive samples,a prevalence of 39%for vv strains and 61%for classical strains was noted.These results indicate that despite the vaccination against IBDV,the vv form of this pathologie continues to cause serious problems for Moroccan broiler chickens.The obtained results indicate the successfully detection of IBDV and differentiated all vvIBDV strains from non-vvIBDV strains;Avian infectious agent RNA viruses tested are negative,demonstrating great specificity of the assay.The results obtained indicate that this method is suitable as a routine laboratory test for the rapid detection and differentiation of IBDV strains in samples of avian origin.

Development of a Microplate Lectin-Capture RT-PCR (MLC-RT-PCR) for the Detection of Avian Infectious Bronchitis Virus

Rapid, sensitive and specific methods are necessary to confirm the diagnosis of outbreaks of avian infectious bronchitis virus (IBV) infection. The amplification of IBV genome by reverse transcription followed by polymerase chain reaction (RT-PCR) has been one of the most used methods for the detection of this virus in clinical samples. To reduce the time and the number of steps in the molecular diagnosis of IBV, we developed a sensitive and rapid detection method based on viral capture by a lectin (Concanavalin A—Con A) in the microplate wells, followed by RT-PCR to amplify the S1 gene. The detection limit of IBV was 103 EID50/ml for the amplification of 5’part of the S1 gene, and 104 EID50/ml for the amplification of full S1 gene. This technique was specific for IBV detection, and no amplified products were detected for other avian viral pathogens (bursal infectious disease virus, avian metapneumovirus and Newcastle disease virus). The MLC-RT-PCR was as sensitive as conventional RT-PCR, and virus isolation method for the detection of IBV in tissue samples collected from experimentally infected birds. The MLC-RT-PCR technique demonstrated a great potential for the rapid and specific diagnosis of IBV.

Comparison of the Serum Proteins and Immune Responses of Velogenic Newcastle Disease Virus Infected Chickens and Ducks

The effect of velogenic Newcastle disease virus (vNDV) on the immune responses and serum proteins was investigated in six-week-old ducks and chickens. Results showed that weight loss was markedly significant (p < 0.05) from days 3 - 21 (PI) in chickens and mild (p < 0.05) on days 3 and 15 PI in ducks. The antibody response obtained showed significant (p < 0.05) increase in infected chickens (IC) than those of the infected ducks (ID). While the total serum protein and serum globulin increased significantly (p < 0.05) in IC on days 7 and 14 PI, they decreased significantly (p < 0.05) in ID only on day 21 PI. The immune responses and serum protein values in this experiment X-ray showed less susceptibility of ducks when compared with the chickens. This may be related to marked anorexia and severe dehydration observed in the latter consequent upon serum concentration. Ducks could be maintaining the endemicity of Newcastle disease (ND) as reservoir host.