Comparative studies of serum neuraminidase, free and erythrocytes surface sialic acid, packed cell volume and haemagglutination inhibition antibodies of chickens vaccinated with different Newcastle disease virus vaccines

Comparative studies of serum neuraminidase, free and erythrocytes surface sialic acid, packed cell volume and haemagglutination inhibition antibody titres were carried out on a total of seventy-five Shaver Brown chickens that were vaccinated with different Newcastle disease virus (NDV) vaccines. Trial birds were primed with NDV Hitchner B1 vaccine prior to NDV La Sota and NDV Komarov vaccines administration. Twenty-five chickens received NDV La Sota vaccine; a further 25 were vaccinated with NDV Komarov vaccine, while 25 chickens served as control. The values of neuraminidase activity (NA), free serum sialic acid (FSSA), erythrocytes surface sialic acid (ESSA) concentration, packed cell volume (PCV) and haemagglutination inhibition (HI) antibody titres were determined sequentially for each chicken. The highest daily mean HI antibody titres of log2 7.12 ± 1.20 and log2 8.98 ± 1.73 were obtained from chickens vaccinated with NDV La Sota vaccine and NDV Komarov vaccine, respectively, by day 15 post-vaccination (pv). Neuraminidase activity and FSSA concentration increased gradually from days 1 to 5 pv in the two groups of chickens. In general, the period of increased NA and FSSA concentration coincided with the time of a decline in values of ESSA and PCV in the two vaccinated groups of chickens. It was concluded that chickens which received NDV Komarov vaccine had higher daily mean values of NA, FSSA and HI antibody titres than their counterparts which received NDV La Sota vaccine

Učinak nekih inhibitora na neuraminidazu soja Kudu 113 virusa newcastleske bolesti.

A total of two hundred samples from dialysed neuraminidase of Newcastle disease virus (NDV) Kudu 113 strain were used for the experiment. Fifty samples each were used for various concentrations of paranitrophenyl oxamic acid (PNPO), salicyl oxamic acid (SOA), silver nitrate (AgNO3) and ethylene diamine tetra acetic acid (EDTA), to test their inhibition effects on the neuraminidase activity of NDV Kudu 113 strain in vitro, using thiobarbituric acid assay method. Of these inhibitors, AgNO3 had the highest inhibition effect on the neuraminidase activity of NDV Kudu 113 strain. Silver nitrate reduced the activity of this enzyme from 140.24 μMol/min for enzyme without AgNO3 to 32.36 μMol/min when AgNO3 was added to the enzyme at a concentration of 0.47 M. Paranitrophenyl oxamic acid had the least inhibition effect on the neuraminidase activity of NDV Kudu 113 strain at a concentration of 0.12 M. This inhibitor reduced the activity of the enzyme from 97.08 μMol/min for enzyme without PNPO to 91.70 μMol/min when PNPO was added to the enzyme at a concentration of 0.12 M. Ethylene diamine tetra acetic acid and SOA also greatly inhibited the activity of this enzyme. Salicyl oxamic acid reduced the activity of NDV Kudu 113 strain neuraminidase from 140.24 μMol/min for enzyme without SOA to 59.33 μMol/min when SOA was added at a concentration of 0.47 M. Similarly, EDTA reduced the activity of this enzyme from 124.06 μMol/min forenzyme without EDTA to 75.51 μMol/min when EDTA was added to the enzyme at a concentration of 0.35 M. In all the inhibitors examined, it was observed that their effects on the activity of NDV Kudu 113 strain neuraminidase increased as the concentrations of the inhibitors increased. It was concluded that PNPO, SOA, EDTA and AgNO3 inhibited the neuraminidase activity of NDV Kudu 113 strain in vitro. Therefore, further studies are needed to see if these inhibitors could be of use, after proper clinical trials, as potential agents for reducing Newcastle disease (ND) infections in poultry.Iz soja Kudu 113 virusa newcastleske bolesti pripremljeno je 200 uzoraka dijalizirane neuraminidaze u svrhu istraživanja inhibicijskog učinka različitih koncentracija paranitrofeniloksaminske kiseline, saliciloksaminske kiseline, srebrnog nitrata i etilendiamintetraoctene kiseline. Učinak svake navedene tvari istražen je na 50 uzoraka neuraminidaze postupkom koji se izvodi pomoću tiobarbiturne kiseline. Najveći inhibicijski učinak na aktivnost neuraminidaze imao je srebrni nitrat. On je u 0,47 M koncentraciji smanjio učinak enzima sa 140,24 μM/min na 32,36 μM/min. Para-nitrofeniloksaminska kiselina imala je najslabiji inhibicijski učinak na neuraminidaznu aktivnost pretraživanog soja. Ona je u 0,12 M koncentraciji smanjila aktivnost neuraminidaze s 97,08 μM/min na 91,70 μM/min. Za etilendiamintetraoctenu kiselinu i saliciloksaminsku kiselinu utvrđena je visoka inhibicijska sposobnost. Saliciloksaminska kiselina u 0,47 M koncentraciji smanjila je aktivnost neuraminidaze sa 140,24 μM/min na 59,33 μM/min. Etilendiamintetraoctena kiselina u 0,35 M koncentraciji smanjila je aktivnost enzima sa 124,06 μM/min na 75,51 μM/min. Inhibicijska aktivnost svih enzima povećavala se s njihovom koncentracijom. Zaključuje se da su sve rabljene tvari in vitro zakočile neuraminidaznu aktivnost soja Kudu 113 virusa newcastleske bolesti. Potrebna su daljnja istraživanja za njihovu moguću praktičnu primjenu u sprečavanju newcastleske bolesti

Uloga sijalične kiseline na površini eritrocita u nastanku anemije u jaraca Savannah smeđe pasmine.

Erythrocyte surface sialic acid, free serum sialic acid concentrations and sialidase activity were determined during experimental Trypanosoma evansi infection in Savannah Brown bucks. All infected bucks developed trypanosomosis, with significant decreases in mean packed cell volume to as low as 19.50 ± 2.12% occurring at day 33 post-infection and was significantly lower than the control value of 26.75 ± 0.96. Mean haemoglobin concentrations also declined in the infected bucks with a marked drop of 6.50 ± 0.70 g/dL on day 33 post-infection and was significantly different (P<0.05) from the uninfected (control) group (8.53 ± 0.46 g/dL). The anaemia was preceded by a gradual decline in mean erythrocyte surface sialic acid concentrations occurring 5 days post-infection. There were significant differences (P<0.05) in mean erythrocyte surface sialic acid between the infected and control groups on day 5 and between days 17 to 27 post-infection. A significant (P<0.05) increase in free serum sialic acid concentrations was observed on days 15, 17 and 27 when compared to the control group. There was a significant (P<0.05) increase in the activity of sialidase on days 7, 9, 21, 23, 27, 33 and 37 post-infection.The anaemia caused during infection may be attributable to the activities of the circulating trypanosomes, which produce sialidase (neuraminidase) that resulted in the cleaving off of erythrocyte surface sialic acid, rendering such red blood cells more prone to phagocytosis in the reticuloendothelial system.U jaraca Savannah smeđe pasmine, u tijeku pokusne zaraze tripanosomom Trypanosoma evansi, određivana je koncentracija sijalične kiseline na površini eritrocita, zatim koncentracija slobodne sijalične kiseline u serumu te aktivnost sijalidaze. Svi invadirani mužjaci oboljeli su od tripanosomoze sa značajnim opadanjem srednje vrijednosti hematokrita do vrijednosti 19,50 ± 2,12%. Navedeni je pad uslijedio 33 dana nakon infekcije i bio statistički značajan u odnosu na kontrolnu vrijednost 25,75 ± 0,96. Srednja koncentracija hemoglobina također je pala u invadiranih mužjaka s najizraženijim padom od 6,50 ± 0,70 g/dL 33. dana nakon zaraze. Ta vrijednost bila je također značajno različita (P<0,05) u odnosu na kontrolnu skupinu jaraca (8,53 ± 0,46 g/dL). Petoga dana nakon infekcije, anemija je nastavljena postupnim opadanjem srednje koncentracije sijalične kiseline na površini eritrocita. Utvrđene su značajne razlike (P<0,05) u srednjoj koncentraciji sijalične kiseline na površini eritrocita između zaražene i kontrolne skupine 5. dana, te između 17. i 27. dana nakon infekcije. U usporedbi s kontrolnom skupinom, u zaraženih životinja utvrđen je signifikantan (P<0,05) porast koncentracije slobodne sijalične kiseline u serumu 15., 17. i 27. dana. Aktivnost sijalidaze bila je značajno (P<0,05) povišena 7., 9., 21., 23., 27., 33. i 37. dana nakon infekcije. Pojava anemije tijekom infekcije može se povezati s aktivnošću tripanosoma u krvnom optjecaju koji tvore sijalidazu (neuraminidazu), a time i cijepanje sijalične kiseline na površini eritrocita čineći ih podložnijima za fagocitozu u retikuloendotelijalnom sustavu

In vitro proizvodnja neuraminidaze bakterije Clostridium chauvoei soja Jakari.

The production of neuraminidase (sialidase, EC 3.2.1.18) by Clostridium chauvoei (Jakari strain) was investigated and a novel neuraminidase was detected in the bacterium in vitro using a fluorescent substrate. Neuraminidase activity was reproducible and had a linear relationship with the amount of bacteria at lower bacterial concentrations. The relationship was no longer linear at higher bacterial concentrations (3.33×107 to 4.44×107 cfu/ml). The possible role of this enzyme in the pathogenesis of blackleg disease is discussed. It is concluded that the enzyme could be playing the role of spreading the disease in the tissues of infected ruminants. Therefore, the use of neuraminidase inhibitors to manage the disease clinically should be thoroughly investigated.Dokazana je proizvodnja neuraminidaze (sialidaze, EC 3.2.1.18) bakterije Clostridium chauvoei soja Jakari in vitro pomoću fluorescentnog supstrata. Aktivnost neuraminidaze bila je reproducibilna uz linearni odnos količine bakterija pri nižim bakterijskim koncentracijama. Taj omjer nije bio linearan pri višim bakterijskim koncentracijama (3,33 x 107 do 4,44 x 107 cfu/ml). Raspravlja se o mogućoj ulozi ovog enzima u patogenezi šuštavca. Zaključeno je da bi enzim mogao imati ulogu u širenju bolesti kroz tkiva inficiranih preživača. Uporaba inhibitora neuraminidaze u kontroli bolesti trebala bi biti potpunije istražena

Utjecaj godišnjeg doba i spola na ukupan broj stanica, hemoglobin i ukupne proteine u krvi autohtonih golubova u Zariji, sjeverna Nigerija.

Blood samples were obtained from 178 indigenous pigeons, comprising 103 males and 75 females, with the aim of determining the effect of season and sex on their packed cell volume (PCV), haemoglobin (Hb) and total proteins (TP) using routine laboratory procedures. The samples were obtained during the harmattan, hot and rainy seasons. Correlation coefficients between meteorological elements and PCV, Hb as well as TP, were negative during the hot season. The highest PCV and Hb values of 37.50 ± 1.23 (%) and 129.0 ± 03.0 g/L, respectively, were obtained during the rainy season, while the lowest PCV and Hb values of 29.12 ± 0.88 % and 97.2 ± 2.9 g/L, respectively, were obtained during the hot season. The TP values of 78.2 ± 13.0 g/L and 82.5 ± 10.0 g/L obtained during the hot and rainy seasons, respectively, were significantly (P<0.05) higher than the value of 64.5 ± 3.3 g/L obtained during the harmattan season. PCV and Hb values were higher (P<0.01) in male than in female pigeons during the three seasons. The results show seasonal and sex variations in PCV, Hb and TP values of indigenous pigeons. In conclusion, the rainy and harmattan seasons are apparently more conducive to the rearing of pigeons than the hot season in the Northern Guinea Savannah zone of Nigeria.Istraživanje je provedeno na 178 autohtonih golubova, a među njima 103 mužjaka i 75 ženki, kako bi se odredio utjecaj godišnjeg doba i spola na ukupan broj stanica, količinu hemoglobina i ukupnih bjelančevina. Golubovima je vađena krv u doba harmatana, kao i tijekom sušnog i kišnog razdoblja. Istraživanjem je potvrđen negativni korelacijski koeficijent između meteoroloških uvjeta i ukupnog broja stanica u krvi, hemoglobina i ukupnih bjelančevina tijekom sušnog razdoblja. Najveći broj stanica (37 ± 1,23%) te vrijednosti hemoglobina (129,0 ± 3,0 g/L) je dokazan u kišnom razdoblju. Najmanji broj stanica (29,12 ± 0,88%) kao i vrijednosti hemoglobina (97,2 ± 2,9 g/L) utvrđen je u tijeku sušnog razdoblja. Vrijednosti ukupnih bjelančevina (78,2 ± 13,0 g/L) u tijeku sušna razdoblja te 82,5 ± 10,0 g/L tijekom kišnog razdoblja bile su značajno različite (P<0,05) u odnosu na vrijednost (64,5 ± 3,3 g/L) utvrđenu u tijeku razdoblja harmatana. Ukupan broj stanica i količina hemoglobina bili su veći (P<0,01) u mužjaka tijekom svih razdoblja. Analizom dobivenih rezultata može se zaključiti da je kišno razdoblje kao i razdoblje harmatana mnogo pogodnije za uzgoja golubova nego sušno razdoblje na području sjeverne gvinejske savanske zone Nigerije

Effect of Extract (Interface) from Stem Bark of Antidiabetic Anogeissus leiocarpus (African Birch Tree) on Random Blood Glucose Levels of Adult Female Wistar Rats: Optimisation for Therapeutic Hypoglycaemic Dose

The LD50&nbsp;of the interface portion of ethyl acetate and n-hexane extract from antidiabetic Anogeissus leiocarpus stem bark was greater than 2000mg/kg.bd.wt., with a wide range of safety value. In addition it caused no death 24hours and thereafter and had no effects on pregnancy, gestation, parturition, reproductive performance with no teratogenic effects on pups of Wistar-rats. This led to investigation of the effects of the extract on ovarian functions of adult female Wistar-rats but strategically optimized to obtain appropriate therapeutic hypoglycaemic-dose. On day 0, 20 adult-female Wistar-rats weighing between 155 and 235 g were separated, at random, into 4 cages (A-D) of 5 rats each, unadministered A (normal control), administered B (5 mg/kg.bd.wt.), C (10 mg/kg.bd.wt.) and D (20 mg/kg.bd.wt.); to each group, an adult male Wistar-rat was added. They were on feeds (superstarter chow) and water ad libitum. Groups B and C were reduced to 4 rats each due to exclusion criteria. On day0, baseline parameters, were collected from all rats. Thereafter, from day0 to day12, each female rat received the optimized dose of the interface and samples were collected for random blood glucose (RBG), vaginal cytology and assay for oestrogen and progesterone. Blood samples were collected after day 12 for clinical biochemistry analyses. At day 1, mean RBG values were significantly (P&lt;0.05) higher (129.5±57.7 mg/dl) in group B than values of groups A,C&amp;D which were 99.0±13.2, 101.8±3.2 and 102.0±11.9 mg/dl, respectively. From days 2 to 12, the mean RBG values significantly (P&lt;0.05) decreased to 98.3±15.6 in group B. The mean bodyweights of group B decreased from day 2 to14. In group D, mean values of body weights equally decreased on day14, but after increase on day7 Conclusion: Daily administration of 5 mg/kg/bd.wt. of the extract (interface) of A. leiocarpus stem-bark is therapeutically hypoglycaemic as it reduced RBG significantly (P&lt;0.05) up to day12. The clinical application of lower doses is suggested to maintain normoglycaemia for a while after “crashing” down the hyperglycaemia of DM with a much higher dose. The shelf-life of extract (interface) of A. leiocarpus stem bark is greater than 17 months, when stored at room temperatur

Acute and Delayed Oral Toxicity Studies and Observations on Pregnancy, Gestation and Reproductive Performance of Wistar Rats Administered Limit Dose of Purified Extract from Stem Bark of Antidiabetic Anogeissus leiocarpus (African Birch Tree)

Objectives: The maximum purification of the extracts of antidiabetic Anogeissus leiocarpus stem bark led to a quest for its lethal dose (LD50) and effects on pregnancy, gestation, pups (kitten) and reproductive performance of Wistar rats. Design: Extracts of A. leiocarpus were subjected to column and thin layer chromatography until final purification. Five (5) each of adult male and female rats were used for the study. The revised limit test dose of 2000mg/kg.bd.wt. was used to evaluate the acute oral toxicity and effects on reproductive characteristics of the purified extracts on Wistar rats. The rats were monitored for instant death and 24hours later, 3 each of male and female rats were humanely sacrificed. On day 0 and 24hours later biomarkers of liver damage, AST, ALT, and ALP and that of kidney, urea, were assayed. The remaining 4 rats (2 males and 2 females) were monitored for additional 14days and thereafter for any effect of the limit dose on reproductive activities. Results: The limit dose caused no death within the first 24hours and no hepato-renal damages. Pregnancy, gestation, parturition, pups (kitten) and reproductive performance were normal. The median lethal Dose (LD50) is &gt; 2000mg/kg./bd.wt. Conclusion: Animal experimentation, using Wistar rats showed that purified extracts of A. leiocarpus stem bark as a non-conventional drug for diabetes mellitus, has no effects on pregnancy and reproductive performance. There was no teratogenic effect on pups. The compound has a wide range of safety value. Keywords: Oral Toxicity, Pregnancy, Gestation, Reproductive Performance, Wistar Rats, Anogeissus leiocarpus, Purified Extrac