Sustainable biosynthesis of lycopene by using evolutionary adaptive recombinant Escherichia coli from orange peel waste.

This study aimed to evaluate the hydrolysates from orange peel waste (OPW) as the low-cost carbon source for lycopene production. Initially, the dilute acid pretreatment combined with enzymatic hydrolysis of OPW resulted in a total sugar concentration of 62.18 g/L. Meanwhile, a four-month adaptive laboratory evolution (ALE) experiment using a d-galacturonic acid minimal medium resulted in an improvement in the growth rate of our previously engineered Escherichia coli strain for lycopene production. After evolutionary adaptation, response surface methodology (RSM) was adapted to optimize the medium composition in fermentation. The results obtained from RSM analysis revealed that the 5.53 % carbon source of orange peel hydrolysate (OPH), 6.57 g/L nitrogen source, and 30 °C temperature boosted lycopene production in the final strain. Subsequently, the optimized treatment for lycopene fermentation was then conducted in a 5 L batch fermenter under the surveillance of a kinetic model that uses the Logistic equation for strain growth (µm = 0.441 h-1), and Luedeking-Piret equations for lycopene production (Pm = 1043 mgL-1) with growth rate constant (α = 0.1491). At last, lycopene biosynthesized from OPH was extracted and analyzed for qualitative validation. Likewise, its data on phytic acid (between 1.01 % and 0.86 %) and DPPH radical scavenging (between 38.06 % and 29.08 %) highlighted the better antioxidant capacity of lycopene. In conclusion, the OPH can be used as a fermentation feedstock which opens new possibilities of exploiting fruit crop residues for food and pharmaceutical applications.

Serological and Molecular Detection of Bovine Brucellosis at Institutional Livestock Farms in Punjab, Pakistan.

Bovine brucellosis remains a persistent infection in ruminants in Pakistan. A total of 828 (409 buffaloes and 419 cattle) sera were collected from 11 institutional-owned livestock farms in Punjab, Pakistan. The samples were tested by rose bengal plate agglutination test (RBPT) and indirect enzyme-linked immunosorbent assay (iELISA). The seroprevalence along with 95% confidence interval (CI) was determined. Univariable and multivariable analysis of the epidemiological background data was conducted and odds ratio (OR) was calculated to understand any association between the risk factors and the seroprevalence. An overall seroprevalence of 3.9% (Positive/Tested = 32/828) and 3.3% (27/828) was detected by RBPT and iELISA, respectively. The seroprevalence of 5.6% (CI 3.6-8.3) and 4.7%, (CI 2.8-7.2) and the odds ratio of 2.63 (CI 1.20-5.77) and 2.50 (CI 1.08-5.78) for testing positive by RBPT and iELISA, respectively were significantly higher (p < 0.05) in buffaloes than in cattle. Breed, sex, history of abortion and retention of fetal membranes (RFM) in the animals were not found statistically significantly associated with the infection. RBPT and iELISA based results agreed almost perfect (k = 0.877). In total, Brucella abortus-DNA (9/27) was amplified from seropositive samples by real-time polymerase chain reaction. This study identified for the first time the etiological agents of brucellosis at a molecular level at institutional-owned livestock farms in Pakistan.

Serological and Molecular Investigation of Brucella Species in Dogs in Pakistan.

Brucellosis is an important bacterial zoonosis caused by B. abortus and B. melitensis in Pakistan. The status of canine brucellosis caused by B. canis remains obscure. In total, 181 serum samples were collected from stray and working dogs in two different prefectures viz. Faisalabad (n = 87) and Bahawalpur (n = 94). Presence of antibodies against B. canis and B. abortus/B. melitensis was determined using the slow agglutination test (SAT) and ELISA, respectively. Real-time PCR was performed to detect and differentiate Brucella DNA at the species level. In Faisalabad, the serological prevalence was found to be 9.2% (8/87) and 10.3% (9/87) by SAT and ELISA, respectively. Only one of the ELISA positive samples (1.15%) yielded amplification for B. abortus DNA. In Bahawalpur, 63.8% (60/94) samples were found positive by SAT; however, none of the samples was positive by ELISA or by real-time PCR. Location, age (≥1 year) and body condition (weak) were found to be associated with B. canis infection, whereas presence of wounds was found to be associated with B. abortus infection only. These findings point towards a risk of transmission from dog to livestock and humans and vice versa. The study expects to draw the attention of concerned authorities towards infection prevention and animal welfare. This study warrants further epidemiological investigation on brucellosis in pet dogs and their owners. To the best of our knowledge, this is first ever report on B. canis and B. abortus in dogs in Pakistan.

Prevalence of Giardia intestinalis and Hymenolepis nana in Afghan refugee population of Mianwali district, Pakistan.

BACKGROUND: Present study aimed to investigate prevalence of Giardia intestinalis and Hymenolepis nana in Afghan refugees visiting Central Health Unit (CHU), Kot Chandana (Mianwali, Northern Punjab) during two years period (February 2007 to December 2009). METHODS: A total of 687 stool samples were collected from different age groups of both genders. Samples were processed under sterile conditions after gross examination. Microscopic examination was done on same day along with eggs (H. nana), cyst and trophozoites (G. intestinalis) detection after staining. RESULTS: The prevalence of G. intestinalis was significantly higher (x2=59.54, p<0.001) than that of H. nana. Females were found more likely to be infected as compared to males (OR: 1.40, 95% CI=1.03-1.92). Prevalence of both parasites decreased with age and highest prevalence was observed in young individuals belonging to 1-15 years of age group (41.8% and 48.7% respectively for H. nana and G. intestinalis, p<0.001). Abdominal distress (OR: 1.13, 95%CI=0.83-1.53), vomiting (OR: 1.13, 95%CI=1.13-1.81) and rectal prolapse (OR: 4.26, 95%CI=1.38-13.16) were the gastro-intestinal clinical symptoms observed in G. intestinalis. Whereas, bloody diarrhea (OR: 1.56, 95%CI=1.00-2.43) and rectal prolapse (OR: 5.79, 95%CI=1.87-17.91) were associated with H. nana infections. CONCLUSIONS: Intestinal parasitic infections are common among Afghan refugees and serious preventive measures should be implemented to promote the safety and healthy lifestyle of these people.