Antibiofilm Effects of Macrolide Loaded Microneedle Patches: Prospects in Healing Infected Wounds.

AIM: The aim of this study was to fabricate polymeric microneedles, loaded with macrolides (erythromycin, azithromycin), using hyaluronic acid and polyvinyl pyrollidone. METHODS: These microneedles were fabricated using a vacuum micromolding technique. The integrity of the microneedle patches was studied by recording their morphologic features, folding endurance, swelling and micro-piercing. Physicochemical characteristics were studied by differential scanning calorimetry, thermogravimetric analysis and fourier transform infrared spectroscopy. In-vitro drug release, antibiofilm and effect of microneedle patch on wound healing were also studied to confirm the efficacy of the formulations. RESULTS: Formulated patches displayed acceptable folding endurance (>100) and uniform distribution of microneedles (10 × 10) that can penetrate parafilm. Differential scanning calorimetry results depict a decrease in the crystallinity of macrolides following their incorporation in to a polymer matrix. Percentage release of azithromycin and erythromycin from the polymeric patch formulations (over 30 min) was 90% and 63% respectively. Broadly, the zone of bacterial growth inhibition follows the same order for Staphylococcus aureus, Escherichia coli and Salmonella enterica. After 5 days of treatment with azithromycin patches, the wound healing was complete and skin structure (e.g. hair follicles and dermis) was regenerated. CONCLUSION: It was concluded that azithromycin loaded microneedle patches can be used to treat biofilms in the infected wounds.

Vector-borne pathogens in dogs of different regions of Iran and Pakistan.

Canine vector-borne diseases (CVBDs) are highly prevalent in tropical and subtropical countries, mainly due to favorable climate conditions and reduced adoption of preventive measures. This study aimed to provide a comprehensive overview on the prevalence of CVBDs in Iran and Pakistan where limited data are available. Blood samples were collected from 403 dogs from six provinces in Iran and Pakistan to assess the presence of pathogen DNA (i.e., Anaplasma spp., Coxiella burnetii, Ehrlichia spp., Rickettsia spp., Babesia spp., Hepatozoon spp., filarioids, and Leishmania spp.). Sera were also screened by an immunofluorescence antibody test for the detection of antibodies against Leishmania infantum. In total, 46.9% of dogs scored positive to Hepatozoon canis being the most frequently detected (41.4%), followed by Anaplasma platys (6.4%), Ehrlichia canis (3.4%), Rickettsia spp. (2.2%), Babesia vogeli (1.0%), and L. infantum (0.3%). A seroprevalence of 9.6% to anti-L. infantum IgG was also recorded. Data reported herein demonstrate that dogs from Iran and Pakistan are at a high risk of CVBDs, particularly of canine hepatozoonosis. Effective control strategies are advocated for minimizing the risk of infection in animals and humans, also in consideration of the zoonotic potential of some pathogens detected.

An Insight Into COVID-19: A 21st Century Disaster and Its Relation to Immunocompetence and Food Antioxidants.

Coronavirus Disease 2019 (COVID-19) ranks third in terms of fatal coronavirus diseases threatening public health, coming after SARS-CoV (severe acute respiratory syndrome coronavirus), and MERS-CoV (Middle East respiratory syndrome coronavirus). SARS-CoV-2 (severe acute respiratory syndrome coronavirus type 2) causes COVID-19. On January 30, 2020, the World Health Organization (WHO) announced that the current outbreak of COVID-19 is the sixth global health emergency. As of December 3, 2020, 64 million people worldwide have been affected by this malaise, and the global economy has experienced a loss of more than $1 trillion. SARS-CoV-2 is a positive-sense single-stranded RNA virus belonging to the Betacoronavirus genus. The high nucleotide sequence identity of SARS-CoV-2 with the BatCoV RaTG13 genome has indicated that bats could be the possible host of SARS-CoV-2. SARS-CoV-2 penetrates the host cell via binding its spike protein to the angiotensin-converting enzyme 2 (ACE2) receptor, which is similar to the mechanisms of SARS-CoV and MERS-CoV. COVID-19 can spread from person to person via respiratory droplets and airborne and contaminated fomites. Moreover, it poses a significant risk to smokers, the elderly, immunocompromised people, and those with preexisting comorbidities. Two main approaches are used to control viral infections, namely, vaccination, and biosecurity. Studies to analyze the antigenicity and immunogenicity of SARS-CoV-2 vaccine candidates are underway, and few vaccines may be available in the near future. In the current situation, the Human Biosecurity Emergency (HBE) may be the only way to cope effectively with the novel SARS-CoV-2 strain. Here, we summarize current knowledge on the origin of COVID-19 as well as its epidemiological relationship with humans and animals, genomic resemblance, immunopathogenesis, clinical-laboratory signs, diagnosis, control and prevention, and treatment. Moreover, we discuss the interventional effects of various nutrients on COVID-19 in detail. However, multiple possibilities are explored to fight COVID-19, and the greatest efforts targeted toward finding an effective vaccine in the near future. Furthermore, antioxidants, polyphenols, and flavonoids, both synthetic and natural, could play a crucial role in the fight against COVID-19.

Molecular Epidemiology and Prevalence of Theileria lestoquardi and Theileria ovis Infection in Goats Infested With Tick Vectors From Multan, Pakistan.

Caprine theileriosis is a major production problem in regions of the world that rely on goats as a major source of milk, meat, and other means of economic income. Theileria ovis and Theileria lestoquardi are responsible for caprine theileriosis in Pakistan. The present study examined the prevalence of Theileria spp. infection using polymerase chain reaction (PCR) and light microscopy of blood collected from goats from Multan, Pakistan. In addition, the prevalence and identity of the genus of ticks feeding on these goats was determined on specimens collected at the time of blood sampling. The current project hypothesized that Theileria spp. prevalence would be higher in goats infested with ticks than goats without tick infestation. Four hundred and sixty-three blood samples from goats were analyzed, and the prevalence of infection was 16.0% (95% confidence interval [CI] = ±5.96) 74 of 463 by PCR and 5.4% (95% CI = ±7.79) 25 of 463 by microscopic examination. Thirty-six (48.6%, 95% CI = ±4.96) and 30 (40.5%, 95% CI = ±7.10) of 74 samples were positive by PCR for T. ovis and T. lestoquardi, respectively. Eight samples (10.0%, 95% CI = ±9.61) had mixed infections of these Theileria species. Infection was observed significantly (P < 0.05) more often in male 22.8%, 95% CI = ±3.85 (23/101) than in female 14%, 95% CI = ±2.36 (51/362) goats. No statistical (P > 0.05) difference in prevalence was present among the three age groups of ≤1 yr (12.6%, 95% CI = ±6.70), 1-4 yr (16.2%, 95% CI = ±8.24), and ≥4 yr (17.7%, 95% CI = ±11.30) goats examined. Two hundred and one (43.4%, 95% CI = ±7.45) goats were infested with ticks, Rhipicephalus spp. and Haemaphysalis spp., which are capable of transmitting Theileria spp. Tick infestation was found to be a significant (P < 0.05) risk factor for infection. Prevalence of Theileria infection in goats was significantly higher (P < 0.05) in the summer compared with the other three seasons. Data from this study support our hypothesis and demonstrate that both T. ovis and T. lestoquardi are highly prevalent in goats from Multan, Pakistan.

Hepatoprotective and hepatocurative effects of Spilanthes acmella Murr against paracetamol induced hepatotoxicity.

Hepatotoxicity is appreciably escalating health dilemma worldwide and the degree of the problem has encouraged importance in the exploration for hepatotherapeutic agents from plants. In the current research work, the hepatoprotective/hepatocurative activity of methanolic extract of Spilanthes acmella Murr aerial parts in paracetamol induced hepatotoxicity was investigated in rabbits by the analysis of different liver enzymes including ALT, AST, ALP along with histopathological investigations. In first phase of the study, paracetamol toxicated rabbits were treated with extract and standard drug jatepar TM. The hepatotoxicant (paracetamol) significantly increased the levels of aspartate transaminase, alanine transaminase, alkaline phosphatase compared to normal control. Spilanthes acmella Murr at (400 mg/kg) reversed the elevation in the level of ALP, AST and ALT caused by the hepatotoxicants and jeteparR TM (standard) also reversed the deleterious effects of the hepatotoxicants. In second phase of this study, extract of Spilanthes acmella Murr was given to rabbits for ten days then paracetamol was administered in one group and level of liver parameters was paralleled with regular control group and the group that was receiving the extract. It is concluded that methanolic extract of Spilanthes acmella Murr aerial parts possesses hepatocurative and hepatoprotective activity.

Prevalence and risk factors for IgG antibodies to Neospora spp. in three types of equids from Southern Punjab, Pakistan.

Equine hosts suffer from neurological disease, congenital infection, and reproductive problems associated with Neospora spp. infection. We conducted a cross sectional study using sera from 631 equids (324 horses, 218 donkeys and 89 mules) from the southern region of Punjab province, Pakistan to determine the prevalence of antibodies against Neospora spp. in this diverse group of equines. Fisk factors associated with seropositivity were evaluated statistically based on equine type, breed, age, husbandry, breeding methods, and reproductive history. Prevalence of antibodies to Neospora spp. was detected using a commercially available competitive ELISA kit. We detected IgG antibodies to Neospora spp. in 23.3% of the equids with prevalence by host being 16.0% in horses, 32.6% in donkeys and 26.9% in 89 mules. Statistically significant (P < 0.05) differences in prevalence were observed among these hosts. Prevalence ranged in breeds of horses from 3.7% in the Morna breed, 12.4% in breeding stock, 26.8% in draughting stock, to 31.4% in unknown breeds. No significant (P > 0.05) difference in prevalence was noted among age groups. Prevalence was significantly (P < 0.05) higher in female equines with a history of abortion than those with no history of abortion. This is the first report documenting Neospora spp. in equines from Pakistan and it provides evidence that Neospora spp. is associated with abortions in female equines in Pakistan.

Prevalence of Toxoplasma gondii, Neospora caninum, and Sarcocystis Species DNA in the Heart and Breast Muscles of Rock Pigeons (Columbia livia).

Little is known about the prevalence of protozoan parasites in the muscles of rock pigeons (Columbia livia). The muscles from 54 (heart from 45 and breast from 54) rock pigeons were examined for DNA of Toxoplasma gondii, Neospora caninum, and Sarcocystis species using PCR. Twenty-four were female and 30 were males. The birds were part of flocks of pigeons housed at the tombs of saints in Lahore, Pakistan. Birds that died or were euthanized due to poor health were submitted for necropsy at the Department of Parasitology, University of Veterinary and Animal Sciences, Lahore, Pakistan, where DNA isolations and PCR were conducted. Nineteen (35.1%) of the birds were positive for T. gondii DNA. Seven males and 12 females were positive. Breast tissue was always infected in T. gondii positive birds, while the heart was infected in 13 (28.8%) of breast positive birds. Five (9.2%) of the pigeons, 2 males and 3 females, were positive for N. caninum. The distribution of N. caninum DNA was more variable in the muscles of pigeons than T. gondii and was found only in the heart of 1 (female), heart and breast muscle of 2 (male), and only the breast muscle of 2 birds (female). One of the 54 rock pigeons (female) was positive for both T. gondii (heart and breast) and N. caninum (heart only). Two of the positive Neospora caninum amplicons were sequenced and had 97% nucleotide identity with N. caninum isolates. Sarcocystis DNA was not found in any bird. The prevalence of T. gondii in rock pigeons and their predation by cats suggest that they may play an unrecognized role in maintaining environmental contamination with T. gondii oocysts by cats. Our study indicates that rock pigeons are intermediate hosts of N. caninum and this information will aid in understanding the epidemiology of N. caninum.

Report: An ex vivo up-take of levamisole molecules by cestode (Monezia expensa) of goat (Capra hirsa) and its detection through RP-HPLC.

Detection of various molecules of drugs remained a prime issue especially in tissues of animals, humans and in their target parasites. The cestode/tapeworms pose a dilemma because of their weird body composition and uptake pattern of nutrients and medicines especially through absorption by tegument. We selected levamisole; thought to be potent antiparasitic/ani-cestodal drug. The uptake of levamisole (LEV) through cestodeal tissues is studied through HPCL in this paper. High performance liquid chromatography technique has been utilized to know the uptake of levamisole in tissues of cestodes of Goat (Monezia expensa) in small ruminants. The drug was exposed to M. expensa by in vitro till its death or a parasite ceases its movement. The tissue/ part of proglattids of the M. expensa were homogenized with some modifications and levamisole extraction was performed with liquid phase extraction method. The evaporation of solvent was done and the residual cestodal tissues were cleaned by solid phase. After the solid phase extraction method, the recovery of drug, detection and quantification of levamisole from cestodal tissues was determined through Reverse Phase Column High Performance Liquid Chromatography (RP-HPLC). Levamisole (LEV) molecules assay was obtained on a C18 reverse-phase (20um, 6mm x 150mm) column at flow rate of 1ml/min using acetonitrile and ammonium acetate as mobile phase and UV detection was done at 254nm. The development of method of Levamisole (LEV) detection from cestodal tissues by HPLC in vitro samples has been demonstrated first time in Pakistan, which can provide the solution of parasitic control and provide in sight in to the uptake of anti cestodal drugs either against human or livestock parasites.

Cutaneous Leishmaniasis in the Metropolitan City of Multan, Pakistan, a Neglected Tropical Disease.

Cutaneous leishmaniasis is a vector-borne infection caused by flagellated parasitic protozoans in the genus Leishmania. Promastigotes are the infective stage, which are transmitted by the bite of female sand flies of the genera Phlebotomus or Lutzomyia. Three clinical forms of Leishmania infection are recognized in humans, and they are caused by different Leishmania species. They are cutaneous leishmaniasis (CL), mucocutaneous leishmaniasis (ML), and visceral leishmaniasis (VL). Up to 2 million cases of leishmaniasis are reported annually worldwide. In Pakistan, CL is widely distributed in endemic areas. In the current study, a survey was conducted to investigate the occurrence of CL in an endemic area of the metropolitan city of Multan, Pakistan. The material for diagnosis of CL was obtained from 461 patients from 239 clinical laboratories and health care centers in Multan. Of these patients, 42.9% (198 of 461) having positive findings of amastigotes in Giemsa stained tissue impression smears. The distribution of CL skin lesions was highly variable. Most patients (76.2%) had a single lesion. Most CL lesions were found on a single hand (23.7%). The prevalence of two CL lesions per patient was 21.7%, while three or more positive CL lesions were observed in 2.0% of patients. The 198 CL-positive patients ranged in age 10-40 yr old, 101 were males, and 97 were females. Logistic regression analysis of leishmaniasis versus age and gender showed that both the age and gender had a significant (P < 0.05) effect on the occurrence of infection. Outbreaks of CL were reported in 14 of the 68 Union Councils of Multan.

Prevalence of antibodies to Neospora caninum in the serum of camels (Camelus dromedarius) from central Punjab, Pakistan.

Infection with the Apicomplexan parasite Neospora caninum occurs in domestic and wild animals worldwide. This parasite causes significant economic losses in the dairy industry worldwide. The current study was designed to assess the prevalence of N. caninum antibodies in the sera of camels (Camelus dromedarius) from Rakh Mahni, Bhakkar, Pakistan, to better understand the epidemiology of this important cause of bovine abortion in Pakistan. Eighty-one sera samples collected from apparently healthy camels (male = 19, female = 62) were tested for antibodies against N. caninum by using a commercially available competitive Enzyme Linked Immunosorbent assay diagnostic kit. Nine (11.1%) of 81 sera samples were found positive for antibodies to N. caninum. No significant (P > 0.05) difference in seroprevalence was observed between male and female camels or between different breeds. An increasing trend of seropositivity to N. caninum was seen with the age of animals indicating postnatal exposure to N. caninum infection. However, this increase in prevalence was not significant (P > 0.05). However, a significant (P < 0.05) difference in seroprevalence was noted among feeding style of animals, i.e., stall feeding, grazing, and mixed (grazing and stall feeding) feeding style. The prevalence was significantly (P < 0.05) higher in female animals 26.1% (6/23) with abortion history that of animals have had no such history 5.1% (2/39). Presence of antibodies to N. caninum in the sera of camels indicates that further studies are required to evaluate the importance of N. caninum as a cause of abortion or neonatal disease in these animals.

Epidemiological survey of helminths of goats in southern Punjab, Pakistan.

The main purpose of this study was to investigate the prevalence of helminths of goats such as Fasciola hepatica, Haemonchus contortus, Paramphistomum cervi, Oesophagostomum columbian, Cotylophoron cotylophorum, Monezia expansa, Oestertagia oestertagi and Oestertagia circumcincta. The overall prevalence of all species of helminthes was 52% in goat. The study was designed to investigate the factors of helminths prevalence on the basis of sex and age of goat with the help of Chi-square. All the results obtained were non-significant due to some factors which directly affects the prevalence of helminths.