Changes of the Levels of Blood NO and TNF-αConcentrations in Water Buffaloes and Goats Infected with Fasciola hepatica

Changes of concentrations were studied in water buffaloes and goats infected with Fasciola he-patica on blood NO(nitric oxide) and TNF-α(tumor necrosis factor-α). Twenty healthy male castrated water buffaloes of 2 - 3 years old and weighing 300 - 500 kg as well as six goats were confirmed free of fasciolosis by fecal examination and Dot-ELISA. Two studies were conducted using the water buffaloes. In the first experiment, 8 water buffaloes were randomly divided into control group (n = 3) and infection group (n = 5). Each buffalo in the infected group received orally 60 metacercariae of F. hepatica per day for 20 days (total 1 200 metacercariae) to produce a chronic infection. In the second experiment, 12 water buffaloes were randomly divided into infected (n=9) and control group (n = 3). Each buffalo in the infected group was given a single oral dose of 1 600 metacercariae to produce an acute infection. The 6 goats were randomly divided into two infected groups and a control group. The sheep in two infections received a single oral dose of 200 and 500 metacercariae respectively, the control group remained uninfected. Blood NO and TNF-a concentrations of the test animals were measured by a reductive enzyme assay and RIA, respectively. Blood NO concentration in both acutely and chronically infected water buffaloes progressively increased from week 3 post-infection and was significantly greater than that of the control group (P<0. 05) at the 5th week (acute infection) and 7th week (chronic infection), and remaining at higher concentration for the remaining period of the studies. Blood TNF-αconcentrations in both chronically and acutely infected water buffaloes also increased after infection. In the goat experiment, plasma NO concentrations in both infection groups increased from week 3 after infection, and remained higher than that of the control group until the end of the experiment. TNF-α concentrations in goats in infection group 1 and 2 gradually increased after infection and were significantly greater than those of the control group from the 9th to 11th week and from the 11th to 15th week respectively.

Study on Blood Cell Immune Response in Water Buffaloes Infected Acutely with F. hepatica

Action mechanism of blood cell immune response in water buffaloes against acute infection with F. Hepatica was studied. The results showed that after water buffaloes were infected, the total levels of WBC surpassed control group during whole infection period; Eosinophiles (%) of DC were higher than control group at the 2nd week until 19th week, and then dropped and was close to control group; Neutrophiles(%) was low or significantly lower than control group within the 5 - 16th weeks; The total levels of lymphocytes (%) was lower than control group during the whole infection period; T-lymphocytes (%) dropped significantly, but B-lymphocytes(%) had opposite changes from the first week of infection, and they were close to the control group after 11 weeks; RBC-CR] and RBC-IC rosette rates dropped and rose during 2 - 16 and 2-18 weeks, respectively, and then approached the same between both groups. It was suggested that the violent changes of specific and nonspecific immune responses in water buffaloes with acute F. hepatica infection are related with the mechanism against infection with F. hepatica together.

Endocrine and ovarian responses in water buffalo cows immunized against inhibin and subjected to the Ovsynch protocol

The aim of this study was to investigate the feasibility of stimulating ovarian fol icle development in order to improve fertility in water buffalo cows by immunization against inhibin. The experiment was carried out in early summer (May) and included 24 multi-parity crossbred Murrah-Swamp buffaloes that were divided into immunized (n=11) and control (n=13) groups. Each immunized cow was administered with a 2-mL immunogen of mineral oil adjuvant containing 2 mg of recombinant inhibinα-subunit fusion protein. The controls were treated with the adjuvant only. Al animals received Ovsynch protocol treatment, starting on the day of the antigen administration, and they were artiifcial y inseminated upon behavioral estrus. As a result, al of the immunized buffaloes generated antibodies against inhibin during the experimental period and had higher plasma concentrations of fol icle-stimulating hormone (FSH), activin, and estradiol (E2) related to estrous expression. A higher proportion of immunized animals expressed estrus behavior than did the controls (72%vs. 30%, P<0.05). On aver-age, inhibin-immunized buffaloes had signiifcantly more large fol icles (≥9 mm in diameter) than the controls (mean±SEM;1.2±0.1 vs. 0.84±0.1, respectively;P<0.05) and a slightly higher mean total number of fol icles (≥2 mm;11.4±0.7 vs. 9.0±1.1, respectively;P=0.09) and smal (2–4 mm) fol icles (8.81±0.6 vs. 6.84±1.0, respectively;P=0.12). A higher percentage of cows ovulated in the immunized group than in the control group (91%(10/11) vs. 54%(7/13), respectively;P<0.05). Moreover, inhibin-immunized cows had slightly larger corpus luteum (CL) than the controls 9 days after ovulation and signiifcantly higher (P<0.01) post-ovulation peak plasma progesterone (P4) concentrations. Immunization against inhibin also mar-ginal y increased the conception rate 42 days after insemination (45.8%vs. 15.4%;P>0.05). These results demonstrate that immunization against inhibin, coupled with the treatment with the Ovsynch protocol, can constitute a new technique to increase fertility in water buffalo cows.

A comparative pattern of lectin-binding in the endometrial glands of the uterus and placenta of healthy buffaloes and bovines at early gestation

Water buffalo(Bubalus bubalis)and domestic cattle(Bos taurus)are closely related species.However,embryo transfer interspecies has been attempted without any success.The failure in hybrid embryo-implantation is associated with the glycocode in the maternal-fetal interface.Glycosylation patterns have been studied in different species of ruminants;however,in B.bubalis,only the binucleated cells(BNC)have been analyzed.This glycocode is essential for a successful embryo-implantation and can be defined by Lectin-Histochemistry(LHC).The aim of this study is to compare the glycosylation pattern of placenta and uterus in water buffaloes and cattle by LHC.Tissue samples of placenta and uterus from pregnant Mediterranean female water buffaloes(Buf1)and Angus cows(Bov1)were analyzed.All animals were euthanized at 98 days of gestation.LHC was carried out using twelve lectins(Con A,LCA,PSA,sWGA,PHA-e,SBA,UEA-1,WGA,RCA-1,PNA,DBA,BSA-1).The intensity of lectin binding was semiquantitatively scored using a scale of 0(negative)to 3(strongly positive).Difference between species was found in trophoblast layer by PSA,SWGA,PNA and BSA-1,in BNC,and in the mononuclear cells by LCA,PSA,PHA-e,DBA,BSA-1,PNA.In utero,differences in the apical cellular membrane and the secretion of glands were identified by DBA and RCA-1,and in the cytoplasm of those glandular epithelial cells by PHA-e,BSA-1,WGA,and SBA.In both species,BNC presented a strong positive reaction with DBA and SBA,a moderate response by LCA,PHA-e,BSA-1 and PNA lectin,and a low reaction by PSA,UEA-1,SWGA,WGA,Con A and RCA-1.The results found in this study suggest that although both species are closely related,glycosylation patterns in the placenta and uterus are different,thus providing a possible reason for embryo transfer not being possible between these species.