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Back to Journals » Infection and Drug Resistance » Volume 17
pneumoniae in various recreational settings
especially regarding multi-drug resistant (MDR) strains
This superbug is linked to the rapid spread of plasmids carrying these resistance genes
The objective of this study was to evaluate the spatiotemporal prevalence of MDR-K
Southwestern Uganda.Methods: A laboratory-based descriptive longitudinal study was conducted between May and July 2023
we collected eighty water samples in the morning and evening from the hot spring
The temperature at each point was measured prior to sample collection
and two samples were obtained at varying depths
5 mL of each homogenized sample were pre-enriched in brain heart infusion broth
and subsequently in both blood and violet red bile agar
The Kirby-Bauer disk diffusion method was performed
followed by the detection of carbapenemase (CR) and extended-spectrum β-lactamase (ESBL) production
Polymerase chain reaction showed resistance genes viz
Data were analyzed using SPSS-20 to obtain chi-square tests and regression analysis.Results: K
pneumoniae accounted for 30.0% of isolates obtained from Kitagata hot springs
with all isolates classified as multi-drug resistant
95.8% of isolates harbored blaTEM gene alone and both blaTEM and blaCTX genes
with 25% harboring all three resistance genes
pneumoniae had a higher prevalence (35.0%) compared to the wet season (25.0%)
pneumoniae significantly increased over the course of the study
The presence of the three studied resistance genes in the isolates showed a positive correlation with the second phase of sample collection and the dry season but exhibited a negative correlation with temperature
except for isolates harboring either blaTEM alone or blaTEM+KPC+CTX genes.Conclusion: Kitagata hot spring serves as a hotspot for continuous dissemination and acquisition of MDR-K
pneumoniae harboring resistance genes that encode for ESBL and CR production
The healthcare sector ought to implement an ongoing monitoring and surveillance system as well as robust antimicrobial resistance stewardship programs aimed at delivering health education to the community.Keywords: antimicrobial resistomes
this study highlighted the high rate of intercontinental
and local transmission of these resistance genes by visitors or tourists
Kitagata hot springs continue to be significant locations for the potential spread and acquisition of K
pneumoniae superbugs among tourists and individuals with severe illnesses who partake in bathing in the geothermally heated groundwater
Additionally, the increasing prevalence of AMR bacteria in hot springs located in developing nations may stem from poor sewage management, unsanitary environments, and insufficient hygiene practices.18 This ultimately leads to heightened morbidity and extended hospital stays, potentially accompanied by elevated mortality rates within the human population.19
there is insufficient information available regarding the occurrence and spatiotemporal prevalence of MDR-K
this study is necessary to address this knowledge gap
This study was a descriptive longitudinal laboratory investigation that collected quantitative data to evaluate the spatiotemporal prevalence of MDR-K
The study was conducted between May and July 2023
Figure 1 Map of Sheema district showing the study area (Kitagata hot spring)
The water samples were analyzed at the Microbiology Laboratory of the Kampala International University campus and teaching hospital at KIU
pneumoniae across various points of the hot spring during different climatic seasons
two samples (surface and ground/sediment water) were collected in the early morning (0600 hrs) and evening (1900 hrs)
twice within each climatic period (dry and rainy seasons)
with a one-week interval between collections
As described above, approximately 300 mL of water was collected in sterile bottles (Nalgene, New York, NY, USA) from each of the designated sampling sites. After collection, samples were placed in a cooler box (4–8°C) and then transported to the laboratory within 1 h.22 The samples were processed for analysis upon arrival in the laboratory
The water temperature at the designated sampling points was assessed in triplicates just before sample collection using a digital thermometer
The presumptive colonies of K. pneumonia were subjected to preliminary conventional biochemical tests, such as negative tests for indole, methyl red, and oxidase and positive tests for Voges-Proskauer, Citrate, catalase, triple sugar iron fermentation, and delayed urease positivity, as presented in Table S1.3,24
The inoculated plates were allowed to air-dry at room temperature for 15 minutes
sterile forceps were used to place the chosen antibiotic discs (Oxoid
UK) onto the surface of the inoculated agar plates
The antibiotics included: Amoxycillin-clavulanic acid (AMC
Sulphamethoxazole-trimethoprim (SXT 30 μg)
Meropenem (MRP10 μg) and Erythromycin (E 15 μg)
the plates were aerobically incubated overnight at 37 °C
Zones of inhibition were measured (in millimeters) and thereafter interpreted using CLSI standards. The standard MDR strain of K. pneumoniae ATCC BAA-1705 was adopted as the positive control.7 All isolates that exhibited multi-drug resistance were considered for genotypic characterization of resistance genes (blaCTX-M
Table 1 Genes and Their Respective Primers
The PCR master mix was prepared as follows: 12.5µL Hot Start Taq2x master mix (M0496S)-New England Bio-labs
1.0µL reverse primer (10µM) for blaCTX-M and blaTEM genes while for blaKPC gene
5.0µL DNA template and 5.5µL RNAase-Free-H2O
A conventional PCR Thermocycler (CLASSIC K960 Thermal Cycler) was used for PCR amplification. The program was set with initial denaturation at 95 °C for 3 min, followed by 40 cycles (denaturation at 95 °C for 30s, annealing at (59°C for blaTEM, 55 °C for blaCTX-M and blaKPC genes) for 30s, and elongation at 72 °C for 1 min and a final extension cycle at 72 °C for 5 min.28–30
The gel electrophoresis of the PCR amplicons was conducted using the methods described by Tuhamize et al.30 DNA Amplicon was electrophoresed using 1.5% agarose gel
5µL Safe View ClassicTM DNA stain (cat # G108)
6x loading dye (Thermo Scientific #R0611)
and DNA ladder/marker 1kb (NEB-Biolabs #N3231L)
Electrophoresis was run at 200V and 80mA for 1 h
Bands were visualized using a Gene-Flash Trans-illuminator
The PCR products for the reference strains E
pneumoniae NCTC 13368 were used as positive controls to detect blaCTX-M
pneumoniae ATCC® 700603™ was used as a negative control
The formation of bands at a specific bp in reference to the DNA ladder and positive control was considered positive for a specific gene
whereas those without bands were regarded as negative
The data was inputted into Microsoft Excel and then transferred to the Statistical Package for Social Sciences (SPSS) Version 21 for analysis
Descriptive statistics for the occurrence of K
pneumoniae during the study period were obtained using cross-tabulation
and a chi-square test comparison was conducted
Regression analysis was utilized to calculate Pearson’s product-moment correlation coefficient to determine the relationship between study variables
A significant difference of p ≤ 0.05 was applied for all compared variables
The findings from this study are depicted using graphs
and images (especially for gel electrophoresis)
Table 2 Antibiotic Susceptibility Pattern of K
Pneumoniae Isolated in Kitagata Hot Spring
Figure 3 Gel electrophoresis of PCR products for BlaCTX-M gene from selected K
L- DNA ladder (100 bp to 1000 bp); 1–5 are positive isolates for BlaCTX-M gene (551 bp); N- Non template control; P- Positive control (K
Figure 4 Gel electrophoresis of PCR products for BlaTEM gene from selected K
1–23 are isolates positive for BlaTEM gene (800 bp); N- Non template control; P- Positive control (E
Figure 5 Gel electrophoresis of PCR products for BlaKPC gene from selected K
L- DNA ladder (100 bp to 500 bp); 1–8 are positive isolates for BlaKPC gene (150 bp); N- Non-template control; P- Positive control (K
Figure 6 Prevalence of resistance genes in K
pneumoniae isolated from the sampling site
Each block represent one or more resistome(s) harbored by the test organisms
Table 3 Factors Influencing the Spatiotemporal Prevalence of Klebsiella Pneumoniae in the Sampling Sites
Figure 7 Spatiotemporal prevalence of resistance genes associated with K
pneumonia under conditions such as (a) sampling phases
(c) Time of sample collection and (d) Temperature
This study showed no significant difference in the type of resistance genes carried by the isolates obtained during the morning and evening hours of sample collection. The majority of the samples (50.0%) collected during the morning and evening carried both blaTEM and blaCTX genes. However, 50.0% of the isolates collected in the morning harbored blaTEM only compared to those collected in the evening (4.2%) at p-value >0.05 (Figure 7c)
these patients believed that the water helped heal their wounds
inadvertently contributing to the spread of these superbugs
Nonetheless, Anderson et al39 showed increased incidence rates of K
pneumoniae bloodstream infections during the warm season of the year
possibly elucidating the heightened prevalence of this superbug and its resistance genes noted in the dry season
studies have ascertained a greater risk of cross-infection with environmental K
pneumoniae during the warmer seasons compared to the dry season
the correlation between the dissemination of K
pneumoniae and environmental conditions during summer remains ambiguous
pneumoniae clinical isolates in the same hospital
Lubega et al31 reported 100% MDR-K. pneumonia in surgical site infections, which is in agreement with our findings. The above findings indicate a possible vehicle for the dissemination of resistance genes from patients who attend these hospitals and visit the hot spring for spiritual bathes because of their proximity. Likewise, Iramiot et al19 reported high prevalence level of MDR K
at the human–animal interface in Kasese district
This could explain the possible transmission of resistance genes from animals
as they tend to access unprotected hot springs during the night hours
Kitagata hot spring will become a hotspot for the dissemination of infections
posing a risk to all individuals who bathe in the geothermally heated water pool
Our study reported the prevalence of MDR-K
with higher occurrences observed during the dry season compared to the wet season
The dissemination of the pathogen continued to increase over the duration of the study
pneumoniae in hot springs exhibited a significant negative correlation with the temperature at the sampling point
pneumoniae strains were multi-drug resistance
showing particularly high resistance levels to ampicillin
all isolates were identified as ESBL producers
with the majority carrying the blaTEM gene alone or the combination of both blaTEM and blaCTX genes
and 25% harboring all the three resistance genes (blaTEM
The presence of these three studied resistance genes in the isolates was positively associated with the second phase of sample collection and the dry season
but negatively correlated with temperature
except for isolates harboring either blaTEM alone or blaTEM+KPC+CTX together
Kitagata hot springs could become a significant source of infection for individuals bathing in the geothermally heated water pool
this study recommends maintaining cleanliness and implementing basic interventions
to significantly enhance the quality and sustainability of the spring
ensuring clean and safe water for both the community and tourists
The datasets generated and/or analyzed during the current study are not publicly available due [because the isolates were not sequenced; rather
a traditional PCR method was adopted] but are available from the corresponding author upon reasonable request
Ethical clearance for this research was obtained from the Mbarara University of Science and Technology Institutional Research and Ethics Committee and the study was registered with the National Council for Science and Technology (MUST-REC-2023)
We wish to thank all participants of this study
Members of the Department of Medical Microbiology and Immunology
Kampala International University Western Campus
All authors significantly contributed to the reported work
or critically reviewing the article; approved the final version for publication; agreed on the journal for submission; and accepted responsibility for all aspects of the work
Funding for this work was solely by the authors
All authors declare that there are no conflicts of interest in this work
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By Our Reporter
A team of evangelists witnessed residents make a new commitment to Jesus Christ at the famous Kitagata Hot Springs located on the Ishaka–Kagamba Road
This ‘unexpected and miraculous’ moment happened when evangelists and Pastors from Canada’s Harvest City Church invaded the springs traditionally believed to have healing powers – and attract over 1000 visitors per week of which some are tourists
team leader and associate Pastor of Harvest City Church and LIFT Evangelistic Ministries
and Ron Swan who serves as an elder at Harvest City Church
They visited the grounds casually during their ‘day off’ from mission in the district
“When we arrived we were not prepared to see what we saw. The pool was literally formed just by water coming out of the ground and there was about 150 to 200 people sitting in the waters,” Pr. Joel Wells recounted his experience at Kitagata Hot Springs in an update
“Almost all of them had some kind of sickness
deformity or disease and were not wearing much for clothes if any at all
It was a very primitive situation and I immediately felt very uncomfortable
My first reaction was that we didn’t belong there and I told our interpreter how I felt
I felt like we were intruding on something very personal and it didn’t seem right to show up with our cameras and take pictures of this situation
Jeff Barnhardt kept saying to me that we should preach the gospel to these people and pray for the sick
I just wanted to leave because I didn’t feel comfortable
but Jeff kept insisting so finally I turned to him and said
why don’t you preach the gospel?” To which he replied
Jeff was standing on a rock overlooking the pool and with the help of the interpreter he preached a short gospel message,” Pr Joel said
This hot spring locally known as Ekitagata is said to be having its sources from the invisible banks of Nile waters sources that flows through Semuliki in the mountains of the Rwenzori
Traditional healers also visit the venue claiming it has ceasing powers
Jeff Barnhardt’s decision to share God’s word saw many locals turn to God
“When he led [them through] a prayer of salvation
almost everyone in the pool responded and joined in prayer
Then he invited them out of the pool to receive prayer for healing
“we can’t invite a bunch of half-naked people out of the pool for prayer” and so I yelled at Jeff
“Just pray for them as a group while they sit in the pool.” But it was too late
the people were already coming out for prayer,” Pr Joel explained
I stood on the side and watched as Jeff and other members of our team began to pray
spines were being straightened and miracles were exploding
I couldn’t stay on the side lines any longer
One woman approached me and two other team members dragging her leg behind her
It wouldn’t work properly and she was totally lame
the power of God hit her and she began to shake violently
“My fellow team member (Ron Swan) had his hand on her withered leg
and the muscle literally grew under his hand
We were so excited to see her walk away on two strong legs
we would have left and missed what God wanted us to do
We thought we were going to have a day off
but God had a mission for us to accomplish
I’m so thankful for team members who are persistent in their boldness,” Pr Joel said
Joel noted that God uses anyone who is available and willing
He cautioned that sometimes one’s “out of season” is “God’s season.”
All images are courtesy of Lift Evangelistic Ministries
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Uganda | THE INDEPENDENT | Kitagata hot springs in Kitagata town council Sheema district are in a bad shape despite being a destination for hundreds of tourists and residents
The hot springs are adored by local and international tourists who throng the area
While some people visit the springs for adventure
Each visitor pays Shillings 1000 to access the springs
the hot spring is being affected by silting
lack of privacy and inadequate places of convenience
says that run off Water from Ngaro Mwenda stream
which often bursts its banks ends up in the springs
which is used by the visitors is insufficient to handle the large number of guests forcing some of them to uses bushes
He also says a problem of insecurity has cropped as thugs target the property of visitors flocking the area
Bishanga also says they have a challenge of accommodation facilities to carter for the elderly and Persons With disability
He says the available wooden structures are already rotting away and some are in a very bad shape and unfit for habitation
a resident of Kitagata says the biggest challenge has been the failure by the town council authorities to develop the place despite collecting revenue from the hot springs
who hails from Rukungiri and often visits the hot springs
says the state of the hot spring is appealing and unfit to be called a tourist destination
He says the irresponsible disposal of waste and absence of key requirements such as clean water and accommodation among others clearly shows that the authorities are milking a cow that they don’t feed
says the leadership is equally concerned about the current state of the hot springs
He says that they are in discussions with the tourism ministry to guide them before money is invested in the area
Mushabe says they are mobilizing resources to provide security at the hot springs to secure tourists and their belongings
which will be followed by the construction of a hostel
doesn’t specify how much has been set aside for the work and the time frame for the works
Tags Kitagata hot springs Nazario Bishanga Ngaro Mwenda stream Sheema district
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Dozens of locals and tourists gather at the Kitagata hotsprings
Residents in Sheema district have strongly opposed a proposal by the local government to start charging users of the popular “Kitagata” hot spring located in the area
Hot spring users were angered after receiving a letter from the district stating that individuals using the natural facility would be required to part with money
“You are requested that by early next year
everybody shall pay Shs 2,000 daily to use Kitagata hot spring,” read the letter by clerk to council which was copied to the Speaker
Chief Administrative Officer and the Town councillor from Kitagata
Kitagata hot springs are located off the Ishaka–Kagamba Road in Sheema South county in Sheema District ,Western Uganda
Two hot springs sit adjacent to each other
one of the springs known as “Ekyomugabe” was used by the former King of Ankole called “Omugabe”
The other hot spring was dubbed “Mulago Hot Spring” as locals believe it has healing powers as well as blessings to the users
The spring waters in the area contain chemicals and varying amounts of minerals like sodium chloride
among others which are known to have medicinal value in them
Several people with different ailments flock to the springs on a daily basis to bathe in the warm water
The water in the springs can warm up to 80°C (176 °F)
It is estimated that about 800 people visit the springs per week
The springs are fed by River Ngaromwenda which flows from Buhweju district
Godfrey Kiwanda Suubi together with officials from the Uganda Tourism Board visited the springs and promised to work with the district to revamp the area and position it as a tourist attraction
Canon David Kabigumira told SoftPower News on Friday that there are indeed plans by government to take over the management of the Kitagata hotsprings since they are situated on government land
He explained that government has already identified a developer who will give the hotsprings a facelift to make it more attractive to tourists
The district’s stand is that Kitagata hotsprings are a natural resource and they should be a source of revenue for the district,” the Chairman said
the management and services including accommodation provided by private individuals at the hotsprings are unregulated and ineffective
“There are people who built houses near the hotsprings to accommodate guests who spend the night
But there are no toilets and the ecosystem has also facilitated insecurity since criminals use these houses as hideouts,” he added
“The hotsprings should be developed as tourist sites
We have already identified some people who will do this
We need to establish bath tabs where people can bathe conveniently
The hotsprings in their current state embarrass some of the visitors
He said that the decision to develop the hotsprings was reached after holding a meeting with local leaders including LC1 and LC2 Chairpersons who agreed to the proposal
He however clarified that the district has not imposed any fee for users of the hotsprings at the moment but said that the town council would soon convene a meeting and come up with a harmonized fee
Asked what the locals stand to benefit from this facelift
Kabigumira said that the district will assess the needs and services required by the community and provide them
some of the residents in the community near the hotsprings are opposed to the idea claiming that the springs are a free God given feature which should not be taxed by anybody in the area
Prof Ephraim Kamuntu who doubles as the Minister for Tourism
Kabigumira are behind the scheme which they say is intended to benefit the two
“Go and tell Kamuntu and Kabigumira that they should leave alone our Kitagata
This hot spring was here even before we were born
if you want to bring your poverty on our hot spring
know that we as people of Sheema we shall beat and lock you in your houses,” a furious local commented
A native of the area who sought to speak on condition of anonymity said the hot spring is a God given resource and that when authorities attempted to impose a tax for its use in the early 1990s
the warm water turned cold and it was covered by papyrus
the water again turned warm and the papyrus disappeared,” the 72-year-old woman told this website
government leaders should refrain from imposing a tax on our Kitagata [hotspring] which was created and blessed by God to cure people’s diseases,” she added
In order to ensure good hygiene of the facilities
locals in the area have over the years established rules to regulate the use of the hotsprings
Users are not allowed to engage in fighting
Children have to be dipped in dressed in their nappies
David Atwiine on his part labelled critics of the move to tax the users of the hotsprings as saboteurs of development
He said authorities at the district level will engage the community in a meeting next week in a bid to reach a common ground
“We need to advise our people such that they can accept to pay that revenue,” Atwiine told SoftPower News
By press time plans to fence the springs as well as constructing a gate on the entrance were underway
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