Characterization of a novel nicotinamide adenine dinucleotide-cytochrome b5 reductase mutation associated with canine hereditary methemoglobinemia

Hereditary methemoglobinemia associated with nicotinamide adenine dinucleotide-cytochrome b5 reductase (b5R) deficiency is a rare autosomal recessive disorder in animals. Recently, nonsynonymous b5R gene (CYB5R3) variants have been reported to be associated with canine and feline hereditary methemoglobinemia. However, the underlying molecular mechanisms of canine and feline methemoglobinemia caused by these nonsynonymous variants have not yet been reported. Previously, we reported a Pomeranian dog family with hereditary methemoglobinemia, carrying CYB5R3 mutation of an A>C transition at codon 194 in exon 7, replacing an isoleucine residue with leucine (p.Ile194Leu). In this study, we investigated the enzymatic and structural properties of the soluble form of wild-type and Ile194Leu canine b5Rs to characterize the effects of this missense mutation. Our results showed that the kinetic properties of the mutant enzyme were not affected by this amino acid substitution. The secondary structure of the wild-type and Ile194Leu b5Rs detected by circular dichroism showed a similar pattern. However, the mutant enzyme exhibited decreased heat stability and increased susceptibility to trypsin hydrolysis. Moreover, the thermostability and unfolding measurements indicated that the mutant enzyme was more sensitive to temperature-dependent denaturation than the wild-type b5R. We concluded from these results that unstable mutant enzyme properties with normal enzymatic activity would be associated with hereditary methemoglobinemia in the Pomeranian dog family.     

Suppressive effect of deoxynivalenol, a Fusarium mycotoxin, on bovine and porcine neutrophil chemiluminescence: an in vitro study

We evaluated the immunotoxicity of deoxynivalenol (DON), a Fusarium mycotoxin, on bovine and porcine neutrophils in vitro by using two function parameters, luminol-dependent chemiluminescence and random migration under agarose. A 2-hr DON treatment suppressed the chemiluminescence of bovine and porcine cells by 42% and 35% (on average) at 10(-5)M, and by 19% and 26% at 10(-6)M. Slight suppression was observed at concentrations lower than 10(-6)M. However, after an 18-hr DON treatment, random migration of neutrophils of both species remained unaffected, even at the highest concentration (10(-5)M). Although further extensive studies are needed, to our knowledge this is the first study to have revealed in vitro that DON can affect neutrophil function.     

Milk utilisation patterns in the low-input production systems in South Africa

Understanding the utilisation patterns of milk assists in designing appropriate dairy development schemes in rural communities. The objective of the study was to determine milk utilisation patterns in different smallholder farming systems in the Eastern Cape Province of South Africa. Data were collected through the administration of recording sheets to 130 randomly selected households in Alice, Fort Beaufort and Queenstown districts. Amounts of milk produced per household ranged from 9 to 21 l per household per day. Milk consumption/household/day was similar among the three districts. Milk consumption in the early lactation doubled the amount consumed in late lactation (P < 0.05). Milk in the communal areas was largely utilised as fresh or sour milk. Fresh milk was mostly used in tea/coffee or to make porridge for children. Sour milk was consumed with thick boiled maize meal. Fort Beaufort (10.2 ± 1.37 l/day) had the highest sour milk sales whilst Queenstown had the highest fresh milk sales (9.7 ± 5.57 l/day). It was concluded that quantities of milk consumed or sold as fresh or sour were generally low and varied across smallholder farming systems.     

Effect of supplementation of Simada sheep with graded levels of concentrate meal on feed intake,digestibility and body-weight parameters

The experiment consisting of 7 days of digestibility and 90 days of feeding trial was conducted at Wogda (Ethiopia) to determine the effect of supplementation of graded levels of concentrate mix (CM) on feed intake, digestibility, and body weight (BW) change in hay-based feeding of Simada sheep. Twenty-yearling Simada sheep with a mean initial BW of 17.9 ± 0.81 kg (mean ± SD) were used in randomized complete block design arranged into five blocks of four animals. The four dietary treatments that consisted of hay alone (T1), hay +150 g dry matter (DM; T2, low), hay +250 g DM (T3, medium), and hay +350 g DM (T4, high) CM were randomly assigned to each sheep within a block. The CM consisted of wheat bran (WB), noug seed (Guizotia abyssinica) meal and safflower (Carthamus tinctorius) seed meal at the ratio of (2:1:1), respectively. Supplementation with T2 and T3 increased (P < 0.001) total DM and organic matter intake than the control treatment. Overall, supplementation improved (P < 0.001) crude protein intake, digestibility, feed conversion efficiency, BW gain, and profitability compared to the control, whereas sheep on the high than the low and medium level of supplementation performed better in these parameters among the supplemented treatments. From the results of this study, T4 is recommended as the best level of supplementation since it resulted in better nutrient utilization, animal performance, and profitability.     

Risk factors associated with occurrence of nematodes in free range pigs in Busia District,Kenya

Nematode infections are a serious constraint to pig production, especially where free range pig keeping is practiced. This study investigated the epidemiology of nematodes in free range pigs in Busia District, Kenya. Three hundred and six pigs from 135 farms were sampled for faeces that were analysed for nematode eggs per gram (EPG) of faeces using the McMaster technique. The nematode eggs were also identified to genus and species based on morphology. A questionnaire on risk factors was also administered to the pig owners. The overall prevalence and mean nematode EPG were 84.2% and 2,355, respectively. The nematode eggs were identified as those belonging to Oesophagostomum spp. (75%), Strongyloides ransomi (37%), Ascaris suum (18%), Metastrongylus spp. (11%), Trichuris suis (7%) and Physocephalus sexalatus (3%). The prevalence of nematodes was positively correlated (p < 0.05) with the amount of rainfall in the division of the pigs' origin (all nematodes except S. ransomi). The prevalence of nematodes was also associated with the age of the pigs. A lower burden of nematodes was associated (p < 0.05) with a history of deworming (A. suum) and the provision of night housing (S. ransomi and Metastrongylus spp.). In conclusion, this study has provided information on nematode infections and the associated risk factors for free range pigs in Busia District, which can be used when implementing integrated control measures.     

Seroprevalence of <Emphasis Type="Italic">Brucella</Emphasis> antibodies in camels in Katsina State,Nigeria

A cross-sectional study was carried out to determine the status of Brucella infection in one-humped (Dromedary) camels in the North and Central senatorial districts of Katsina State, Nigeria. Nine hundred and eighty serum samples from live and slaughtered camels were tested. Modified Rose Bengal plate test (RBPT) and serum agglutination test (SAT) with ethylenediaminetetraacetic acid, (EDTA) were used as screening and standard tests, respectively. The prevalence of Brucella antibodies were 110 (11.2%) and 103 (10.5%) for RBPT and SAT, respectively. Of the 472 and 508 serum samples tested from the herds and abattoirs, respectively, 63 (13.3%) and 47 (9.3%) were positive by RBPT while 62 (13.1%) and 41 (8.1%) were positive by SAT, respectively. Based on the results, it was concluded that Brucella antibodies were present in camels in the study area. Poor management practices and mixing of camels with other species of livestock as well as unrestricted movement of camels were proposed to be the reasons for the prevalence of the disease in the study area. In view of the public health importance of the disease, it is recommended that there is the need to develop a strategic plan to decrease spread of brucellosis in the study area.     

Lack of glucokinase regulatory protein expression may contribute to low glucokinase activity in feline liver

In most mammals, glucokinase (GK) acts as a hepatic “glucose sensor” that permits hepatic metabolism to respond appropriately to changes in plasma glucose concentrations. GK activity is potently regulated by the glucokinase regulatory protein (GKRP), which is encoded by the GCKR gene. GKRP binds GK in the nucleus and inhibits its activity. GK becomes active when it is released from GKRP and translocates to the cytosol. Low glucokinase (GK) activity is reported to be a principal feature of feline hepatic carbohydrate metabolism but the molecular pathways that regulate GK activity are not known. This study examined the hypothesis that species-specific differences in GKRP expression parallel the low GK activity observed in feline liver. Hepatic GKRP expression was examined using RT-PCR, immunoblot, and confocal immunomicroscopy. The results show that the GCKR gene is present in the feline genome but GCKR mRNA and the GKRP protein were absent in feline liver. The lack of GKRP expression in feline liver indicates that the low GK activity cannot be the result of GKRP-mediated inhibition of the GK enzyme. However, the absence of the permissive effects of GCKR expression on GK expression and activity may contribute to reduced GK enzyme activity in feline liver. The study results show that the cat is a natural model for GCKR knockout and may be useful to study regulation of GCKR expression and its role in hepatic glucose-sensing and carbohydrate metabolism.     

Effect of spineless cactus intake (<Emphasis Type="Italic">Opuntia ficus-indica</Emphasis>) on blood glucose levels in lactating sows and its impact on feed intake,body weight loss,and weaning-estrus interval

The effect of spineless cactus intake (Opuntia ficus-indica) on blood glucose (BG) levels in lactating sows and its impact on daily and total feed intake (dFI^?1 and TFI, respectively), body weight loss (BWL), and weaning-estrus interval length (WEI) were evaluated. Thirty-four hybrid (Yorkshire × Landrace × Pietrain) sows in lactation phase were used. Sows were divided into two groups: G1 (n = 17) where they received commercial feed and G2 (n = 17) provided with commercial feed plus an average of 2.0 ± 0.5 kg spineless cactus, based on a sow’s body weight. The variables evaluated were BG, dFI^?1, TFI, BWL, and WEI. Statistical analysis was performed by using a fixed and mixed model methodology, under a repeated measurements experiment. Group effects were found on all analyzed variables (P < 0.05). The BG was lower in G2 (55.2 and 64.5 mg/dL pre- and post-prandial, respectively), compared to that in G1 (70.9 and 80.1 mg/dL pre- and post-prandial, respectively) (P < 0.05). G2 showed better performance than G1 for dFI^?1, BWL, and WEI (P < 0.05) whose averages were 5.5 ± 1.8 kg, 7.4 ± 4.5%, and 5.3 ± 1.2 days, respectively. Averages for these variables in G1 were 4.7 ± 1.5 kg, 16.8 ± 4.6%, and 6.1 ± 1.6 days, respectively. Intake of spineless cactus reduced BG levels in lactating sows, generating greater dFI^?1, lower BWL at the end of lactation, and a lower WEI.