1.Physiological and epidemiological study of some parasitic and viral enteric infections in dromedary camels in Al-Muthanna province
Al-Yasari, A.M.R. ; Alsalih, N.J. ; Alsaadawi, M.A.
Tropical Biomedicine 2024;41(No.1):1-13
Understanding the normal physiology of the body is the key to study the changes that occur due to any
infection. It is known that enteric infections play a considerable role in affecting normal body status.
Thus, this study was designed for investigating the enteric infections in Arabian camels in Al-Muthanna
Province. In this investigation, 588 fecal and blood serum samples (for diarrheic camels only) were
collected from the camels in different areas of Al-Muthanna Province, Iraq from both sexes of different
ages during the period from October 2020 up to the end of August 2021. The samples were examined
using routine microscopic examination techniques, hematological techniques, and ELISA for parasitic
and viral identification. Eimeria rajasthani, Isospora orlovi were recorded for the first time in Iraqi
camels with clinical signs of diarrhea, dehydration, and emaciation. The study recorded four types of
protozoa: Eimeria spp., Isospora, Cryptosporidium and Balantidium coli. The recorded types of Eimeria
were E. dromedarii, E. cameli, and E. rajasthani. There was a significant effect of age on infection rates
with Eimeria spp. as the highest Eimeria ratio was in ages of less than two years animals. The infection
rates were also affected with months which reached the highest ratios of Eimeria in October while the
lowest ratio of Eimeria was recorded in July. BVDV infection rate was found in camels that suffered from
diarrhea. There is no significant effect of sex on the onset of the viral disease in camels. For hematological
parameters, there were significant differences in RBCs, WBCs, Hb, and PCV values in protozoal and BVDV
infections. In conclusion, different kinds of protozoal and viral infections were recorded. Some of the
recorded infections were associated with acute clinical signs and have zoonotic importance.
2.Effects of NADPH oxidase inhibitor of the Etlingera elatior (E. elatior) fruits extracts in animal sepsis models
Nurhayatun, E. ; Purwanto, B. ; Ulfia, M.
Tropical Biomedicine 2024;41(No.1):14-19
Infectious diseases with complications of sepsis are still public health concern in both developed and
developing countries. Sepsis is a potentially life-threatening systemic immune response to infection
that can lead to end- stage organ failure and death. Sepsis involves multiple mechanisms such as
neuroendocrine, complement activation, blood coagulation, and fibrinolytic system. Reactive oxygen
species (ROS) is an inflammatory mediator produced by NADPH oxidase activation. This study aimed
to investigate the effects of ethanol extract of E. elatior fruits on NADPH oxidase activity. Forty Mus
musculus mice were randomized divided into five groups (n=8), with the intervention group receiving an
intraperitoneal injection of 0.3 mg/kg BW lipopolysaccharide (LPS). There was a normal group without
LPS injection (N-1), LPS injection only (N-2), and those that received LPS injection and ethanol extracts
of E. elatior fruits containing 2.1 mg/20 g (N-3), 4.2 mg/20 g (N-4), and 8.4 mg/20 g (N-5). NADPH
oxidase activity were measured using ELISA. The oneway ANOVA was used to investigate the differences
between the groups. After administration of the extract at a varied dose, N-5 group the lowest NADPH
oxidase activity (p=0.001). The ethanol extract of E. elatior fruit has antioxidant effects. In this study, a
dose of 8.4 mg/20 g of extract significantly reduced NADPH oxidase activity. The ethanol extract of E.
elatior might be considered a treatment in sepsis.
3.Whole-transcriptome sequencing and bioinformatics analysis of calcified outer-layer tissue from cystic echinococcosis
Wang, S.B. ; Sun, H.H. ; Ma, Y.B. ; Dai, Y. ; Ren, Q. ; Liu, Y.Q. ; Shi, C.H. ; Tao, J. ; Li, J.
Tropical Biomedicine 2024;41(No.1):20-28
Echinococcosis is a common zoonotic disease in livestock; the type with the highest incidence is cystic
echinococcosis (CE). In clinical management, patients with CE of the liver in which the cyst wall is calcified
have been found to have better prognoses than those without calcification. In this study, we collected
calcified and uncalcified cyst wall tissue from patients with hepatic CE and observed significant changes
in the expression of 2336 messenger ribonucleic acids (mRNAs), 178 long noncoding RNAs (lncRNAs),
210 microRNAs (miRNAs), and 33 circular RNAs (circRNAs) using high-throughput sequencing (HTS).
Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of
differentially expressed RNAs (DERNAs: DEmRNAs, DElncRNAs, DEmiRNAs, and DEcircRNAs) were
performed to explore these RNAs’ potential biological functions and signaling pathways. Ultimately,
the results of hematoxylin and eosin (H&E) and terminal deoxynucleotidyl transferase deoxyuridine
triphosphate (dUTP) nick end labeling (TUNEL) staining confirmed the correlation between calcification
and apoptosis of the cyst wall. In summary, this study was an initial exploration of the molecular-biological
mechanism underlying spontaneous calcification of the hydatid cyst wall, and it provides a theoretical
basis for exploring new targets for drug treatment in CE.
4.Isolation and phylogenetic analysis of avian infectious bronchitis virus from an imported chicken meat product in Malaysia
Leow, B.L. ; Shohaimi, S.A. ; Mohd. Yusop, F.F. ; Sidik, M.R. ; Mohd. Saeid, F.H.
Tropical Biomedicine 2024;41(No.1):29-35
Avian infectious bronchitis (IB), a Gammacoronavirus, is a highly contagious upper respiratory disease,
affecting chickens of all ages with a significant economic threat to the poultry industry. In February 2020,
a specimen of imported chicken meat product was received and requested for coronavirus testing. The
result was positive for the avian coronavirus, the IB virus (IBV) by molecular detection in the pre-screening
test. Thus, this study aimed to isolate and characterize the IBV from the specimen. Virus isolation via
egg inoculation was attempted and IBV was successfully isolated. The S1 subunit of the spike (S) gene of
the IBV was amplified, sequenced, and the Basic Local Alignment Search Tool (BLAST) analysis showed
that the IBV has 99% and 98% nucleotide similarity with the Malaysian and China IBVs, respectively. The
phylogenetic analysis indicated that the virus belongs to the GI-19 lineage (also known as the QX strain)
and is grouped with other IBVs from Malaysia and China. The GI-19 lineage is one of the primary IB
strains that circulate in Malaysia. The recovery of the virus may be due to the persistence characteristic
of the virus on meat; and the cold chain practices in the imported food product prolong the survival
of this coronavirus. Though IBV is not identified as a hazard in chicken meat or meat products, raw
food should be cooked thoroughly before being consumed. With the increase in international trade in
poultry and poultry products, disease screening at the entry point and import risk analysis is crucial to
ensure food safety and prevent the introduction of new viruses into Malaysia.
5.Effect of Hypericum thymbrifolium BOISS. ET NOE, Hypericum scabrum L. and Eryngium creticum LAM. plant extracts on Leishmania major, Leishmania tropica and Leishmania infantum/donovani strains and their cytotoxic potential
Ozpinar, H. ; Culha, G. ; Ozpinar, N. ; Kaya, T. ; Kara, B. ; Yucel, H.
Tropical Biomedicine 2024;41(No.1):36-44
Leishmaniasis causes significant morbidity and mortality worldwide. In our country, there has been a
significant increase in the number of cases of leishmaniasis in the last decade. In our study, the effects
of Hypericum thymbrifolium, Hypericum scabrum and Eryngium creticum plant extracts were tested on
Leishmania major, Leishmania tropica and Leishmania infantum/donovani, which were clinically resistant
by not responding to Glucantime® therapy. Cytotoxicity of these extracts were evaluated by XTT method
in the human fibroblast cell line. Possible active ingredients were detected by GC-MS analysis from plant
extracts. Glucantime® resistance was detected at concentrations of 50 µg/mL and lower in 4 of the 7
strains tested. No living leishmania parasites were found in leishmania strains treated with plant extracts
at concentrations of 100 µg/mL or higher. The concentrations of plant extracts included in the study
on the WI-38 human fibroblast cell line were not cytotoxic. According to the GC-MS analysis, several
active substances with biological activities and anti-parasitic effects, such as Thiophene, Germacrene-D,
trans-Geranylgeraniol, Pyridine, and Maleimides, were identified. Based on the findings of the study, it
is believed that these identified active substances when supported by in-vivo studies, will pave the way
for future research and have the potential to be developed as anti-leishmania drugs.
6.Ozone and peroxone disinfection of Toxocara canis eggs in water
Ibá ; ñ ; ez-Cervantes, G. ; Cruz-Bautista, J.D. ; Vargas-De-Leó ; n, C. ; Rojas-Bernabé ; , A. ; Ramí ; rez-Cortina, C.R. ; Nogueda-Torres, B.
Tropical Biomedicine 2024;41(No.1):45-51
Water pollution in developing countries continues to be a major health problem due to various
anthropological activities that contribute to the spread of many parasitic diseases, including those
caused by helminths. The aim of this study is to explore the ability of ozone and peroxone to disinfect
drinking water contaminated samples with Toxocara canis eggs. The oxidants used were ozone and
ozone-hydrogen peroxide combination. The treatment of Toxocara canis eggs was carried out in a 50
ml reactor with an operating volume of 10 ml. The pH conditions (5, 7 and 10) were varied for each
treatment. The treatment effect was calculated by counting eggs and examining the condition of the
larvae larval condition (whole, broken and hatched larvae) using an optical microscope. The experiment
was carried out by exposing the eggs for 60 and 120 minutes to ozone and peroxone. The best results
were obtained for helminths treated with the ozone/hydrogen peroxide combination at pH 10, with an
inactivation of 79.2%. The synergistic effect of ozone combined with hydrogen peroxide allows higher
helminth egg inactivation rates, demonstrating that advanced oxidation processes are a real alternative
to apply in the inactivation of Toxocara canis eggs. The results obtained in this study show that the
ozone and peroxone treatment could be a useful disinfection process to destroy or inactivate Toxocara
canis eggs in processes commonly applied in water treatment.
7.Molecular detection, risk factors, and phylogenetic analysis of tick-borne pathogens in dogs from northern Vietnam
Do, T. ; Bui, K.L. ; Zafar, I. ; Inpankaew, T. ; Galon, M.E. ; Ta, P.A. ; Tran, K.T. ; Hasan, T. ; Shengwei, J. ; Ma, Z. ; Hang, L. ; Amer, M.M. ; Ma, Y. ; Mohanta, K.U. ; El Sayed, A.E.S. ; Xuan, X.
Tropical Biomedicine 2024;41(No.1):52-63
In tropical regions, numerous tick-borne pathogens (TBPs) play a crucial role as causative agents of
infectious diseases in humans and animals. Recently, the population of companion and pet dogs has
significantly increased in Vietnam; however, information on the occurrence of TBPs is still limited. The
objectives of this investigation were to determine the occurrence rate, risk factors, and phylogenetic
characteristics of TBPs in dogs from northern Vietnam. Of 341 blood samples tested by PCR, the total
infection of TBPs was 73.9% (252/341). Babesia vogeli (18SrRNA gene – 30.5%) was detected most
frequently in studied dogs followed by Rickettsia spp. (OmpA gene – 27%), Anaplasma platys (groEL
gene – 22%), Bartonella spp. (16SrRNA – 18.8%), Mycoplasma haemocanis (16SrRNA – 9.4%) and
Hepatozoon canis (18SrRNA gene – 1.2%), respectively. All samples were negative for Ehrlichia canis and
Anaplasma phagocytophylum. Co-infection was detected in 31.4% of the samples (107/341) of which,
A. platys/Bartonella spp. (34/94,10%), Rickettsia spp./B. vogeli (19/94, 5.6%), and M. haemocanis/B.
vogeli (19/94, 5.6%) were recorded as the three most frequent two species of co-infection types.
Statistical analysis revealed a significant correlation between TBP infection and several host variables
regarding age, breed, and living area in the current study. The recent findings reported herein, for the
first time in Vietnam, are essential for local veterinarians when considering the appropriate approaches
for diagnosing these diseases. Furthermore, this data can be used to establish control measures for
future surveillance and prevention strategies against canine TBPs in Vietnam.
8.A colorimetric reverse transcription-loop mediated isothermal amplification (RT-LAMP) method for the rapid detection of SARS-CoV-2 in Thailand
Bhakdeenuan, P. ; Bunchoo, S. ; Klayut, K. ; Srisungngam, S. ; Suphan, O. ; Kongthap, I. ; Suphankong, S. ; Phetsuksiri, B. ; Uppapong, B. ; Rudeeaneksin, J.
Tropical Biomedicine 2024;41(No.1):64-69
COVID-19, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), remains a global
health threat. Timely identification of infected cases is important for appropriate patient management
and the control of viral spread. Simple and cost-effective tests are required to increase access to
testing and early case detection. Here, we describe a colorimetric reverse transcription-loop-mediated
isothermal amplification (RT-LAMP) method to detect SARS-CoV-2. The RT-LAMP could amplify the
orf1ab sequence detectable by visual color change within 45 min at 63 °C. The limit of detection (LoD)
for SARS-CoV-2 RNA was less than 100 copies (13.36) per reaction with no cross-amplification with other
related viruses. Clinical evaluation using leftover RNA samples extracted from 163 nasopharyngeal swab
specimens showed perfect agreement in negative (n = 124) and positive samples with cycle thresholds
(Ct) < 34 cycles (n = 33) detected by real-time reverse transcription-polymerase chain reaction (RT-PCR),
targeting RdRp and N genes as a reference. Overall, the diagnostic accuracy, sensitivity, specificity,
positive and negative predictive values of RT-LAMP in testing were 96.32% (95% CI: 92.16-98.64%),
84.62% (95% CI: 68.47-94.14%), 100% (95% CI: 97.07-100.0%), 100% (95% CI: 89.42-100.0%), and 95.38%
(95% CI: 90.22-98.29), respectively. This RT-LAMP assay is simple and reliable, with the potential to be
an alternative for the rapid detection of SAR-CoV-2 with minimal time and fewer resources compared
to real-time RT-PCR.
9.Interplay of interleukins (IL6, IL10) and 25 hydroxycholecalciferol in asthmatic subjects with chronic post-COVID condition (PCC)
Jaleel, A. ; Namoos, K. ; Asim, S. ; Uppal, S.S. ; Zaman, S. ; Irfan, H. ; Pervaiz, S. ; Tariq, M. ; Shafique, M.
Tropical Biomedicine 2024;41(No.1):70-77
The study aimed to compare and correlate serum levels of IL-6, 10, and 25-hydroxycholecalciferol
in individuals with asthma with and without post-COVID condition (PCC). The study was designed
to investigate the inflammatory response and serum 25-hydroxycholecalciferol status in asthmatics
with and without PCC. A cross-sectional study of 252 subjects (128 asthmatics and 124 non-asthmatic
subjects) was carried out. Interleukins and 25-hydroxycholecalciferol levels were estimated on ELISA.
The principle findings were that IL-6 and 25-hydroxycholecalciferol levels were significantly increased
(p<0.001), while IL-10 levels were non-significant in asthmatics with PCC compared to those without PCC.
However, 25-hydroxycholecalciferol levels were significantly increased, but no significant change was
observed in IL-6, and IL-10 levels in non-asthmatics with and without chronic PCC. A significant positive
correlation (r = 0.258) was found between 25-hydroxycholecalciferol and IL-6 but a significant negative
correlation (r = -0.227) with IL-10 in asthmatics with PCC. Similarly, a significant negative correlation (r
= -0.285) was found between 25-hydroxycholecalciferol and IL-10 but was non-significant with IL-6 in
asthmatics without PCC. The correlation of 25-hydroxycholecalciferol with IL-10 was significant (0.683),
but IL-6 was non-significant in non-asthmatics with PCC. Multiple regression analysis showed that age,
IL-6, gender, and PCC were significantly related in adjusted values to 25-hydroxycholecalciferol. This
study sheds light on the complex liaison between 25-hydroxycholecalciferol levels and inflammatory
responses in asthmatics, especially those with PCC. The findings suggest that although asthmatics with
PCC maintain sufficient levels of 25-hydroxycholecalciferol, they show a substantial increase in the proinflammatory response. This suggests that PCC exacerbates the pro-inflammatory response in asthma.
Moreover, the study reveals that asthmatics, whether with or without PCC, display a negative correlation
between 25-hydroxycholecalciferol and the anti-inflammatory response. This emphasizes the main
influence of asthma on the overall inflammatory response. These findings reveal a complex interplay
between vitamin D levels and inflammatory mediators in asthmatic individuals with and without PCC.
10.Sunlight exposure might account for the relatively low COVID-19 morbidity and mortality in tropical countries
Muhammad, M. ; Loong, S.K. ; Khor, C.S. ; Mohd-Azami, S.N.I. ; Kafle, A. ; Useh, U. ; Bello, B. ; AbuBakar, S.
Tropical Biomedicine 2024;41(No.1):78-83
The present study aimed at exploring whether sunlight exposure might account for the relative
difference in COVID-19-related morbidity and mortality between tropical and non-tropical countries.
A retrospective observational study was designed and data from the World Health Organization
weekly COVID-19 epidemiological update was compiled. We examined the total number of confirmed
COVID-19 cases per 100 000 population, as well as the total number of COVID-19-related mortalities
per 100 000 population. Solar variables data were obtained from the Global Solar Atlas website
(https://globalsolaratlas.info/). These data were analyzed to determine the association of sunlight
exposure to COVID-19-related morbidity and mortality in tropical and non-tropical countries. Results
revealed a statistically significant decrease in the number of confirmed COVID-19 cases per 100 000
population (P<0.001), as well as the number of COVID-19-related mortalities per 100 000 population
(P<0.001) between tropical and non-tropical countries. Analyses of sunlight exposure data found that
specific photovoltaic power output, global horizontal irradiation, diffuse horizontal irradiation and
global tilted irradiation at optimum angle were significantly inversely correlated to COVID-19-related
morbidity and mortality. This suggests that stronger sunlight exposure potentially leads to lower
COVID-19-related morbidity and mortality. Findings from this study suggest that the relatively low
COVID-19-related morbidity and mortality in tropical countries were possibly due to better sunlight
exposure that translates into adequate vitamin D status.


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