Measles Virus:A Childhood Nausiance in Tropical Countries

 Measles isn’t just some childhood nuisance—it’s caused by a virus called Morbillivirus hominis, and it spreads like wildfire. This virus is wrapped in a protective envelope, and its genetic material is a single strand of RNA. Once it gets into your body, usually through the air, it attacks your respiratory system first. But it doesn’t stop there. Soon, it moves through the rest of your body, turning what starts as a cough and fever into something much more serious.

Fig: Measles Virus

Characteristics of Measles virus

Contagiousness: Nine out of ten non-immune individuals will contract it if they come into contact with an infected person, making it one of the most contagious diseases known.

Transmission: Respiratory droplets from coughing or sneezing carry the virus, which is airborne. After an infected person has left, it can remain in the air or on surfaces for up to two hours.

Infectious Period: From four days prior to the rash's appearance until four days following it, people are contagious.

Measles virus cases worldwide in 2025-2026

Country	Number of Cases
Indonesia	14,406
Yemen	9,277
Mongolia	8,483
Pakistan	8,310
India	8,184
Angola	5,823
Nigeria	4,676
Mexico	3,164
Russian Federation	2,939
Laos People's Democratic Republic	2,859
Nepal	690 (2023-2024)

Source: WHO, Measles and Rubella global report, 2026

Modes of Transmission

·         Airborne Spread: When an infected person breathes, talks, coughs, or sneezes, the virus is expelled into the air in small particles.

·         Environmental Persistence: The measles virus can remain active and contagious in the air or on surfaces for up to two hours after an infected person has left the area.

·         Direct Contact: Transmission can occur through direct contact with infected nasal or throat secretions (e.g., kissing, sharing drinks) or by touching a contaminated surface and then touching your own eyes, nose, or mouth.

·         Vertical Transmission: Though rare, a pregnant woman can transmit the virus to her fetus during pregnancy or to the baby during delivery.

Fig: Transmission of measles

Symptoms

1.      First Stage (3–5 Days) 

The illness often starts with cold-like symptoms: 

·         High Fever: up to 104°F (40°C).

·         Cough: Usually dry or "barky".

·         (Runny Nose): Sneezing and nasal congestion.

·         Conjunctivitis: Red, watery, or sore eyes, often with sensitivity to light (photophobia).

·         Koplik Spots: Tiny white or bluish-white spots on a red background inside the mouth or cheeks. 

2.      

Second Stage or Rash Stage

A red, blotchy rash typically appears a few days after the initial symptoms: 

·         Progression: It starts at the hairline or behind the ears and spreads downward to the face, neck, trunk, arms, and eventually the feet.

·         Appearance: Small, flat red spots that may be joined together by raised bumps, forming large blotchy patches.

D     Duration: The rash usually lasts 5 to 7 days before fading in the same order it appeared

 

Fig: Koplik Spots of Face and body of child

Treatment

There is no specific antiviral treatment or cure for measles

Home Care and Symptom Relief

Most people recover at home within 7 to 10 days by managing the following: 

·         Hydration: Drink plenty of fluids (water, broth, or oral rehydration solutions) to prevent dehydration caused by fever or diarrhea.

·         Fever and Pain Management: Use over-the-counter medications like paracetamol (acetaminophen) or ibuprofen to reduce high fever and body aches.

o    Warning: Do not give aspirin to children or teenagers due to the risk of Reye’s syndrome, a rare but life-threatening condition.

·         Eye Care: If eyes are sensitive to light, stay in a darkened room. Use a damp cloth or cooled, boiled water with cotton wool to gently clean crusty or sticky eyes.

·         Rest: Ensure plenty of rest to help the body recover. 

Medical and Specialized Treatments

In more severe cases:

·         Vitamin A Supplements: All children diagnosed with measles should be given two doses of vitamin A supplements given 24 hours apart. This helps prevent eye damage and can reduce the risk of death by up to 50%.

·         Antibiotics: While antibiotics do not treat the measles virus itself, they are prescribed if secondary bacterial infections develop, such as pneumonia or ear infections.

o    Prevention and control

Vaccination (Primary Prevention): The most effective way to prevent measles is through the MMR (measles, mumps, and rubella) or MMRV (includes varicella) vaccine.

Source:MoHP Nepal, 2025.

A joyful mother with her child after the child received the measles-rubella vaccine.

Source: WHO Nepal/S.G.Amatya 

Transport Validation of Microbial Test Samples : A guide for Microbiology Laboratory

Purpose

The Purpose of the study was to establish the evidence that the Microbial sample does not get contaminated during transport after sampling from sampling location to site of analysis (Microbiology Laboratory).

Details of Study

Three different Batches of Raw Material (Microcrystalline Cellulose PH 101) were selected for the Study. The materials were selected based on the basis of scientific judgement that the material serves as excellent growth medium for growth and multiplication of Microorganism if contamination occurs

Sampling System

Sampling for microbial test sample was done in duplicate from two different container of each Control Number using aseptic technique. Sampling was done under RLAF. Sample quantity of approximately 10 gram was done. Sampling was done from three different control number (A, B and C) of Microcrystalline Cellulose PH101 and details of sampling were recorded.

Sampling Preparation, Sampling procedure and Transport Method

a.      Sampling preparation

Sampling vials were cleaned with 1% Labolene solution and rinsed with purified water. After cleaning, the vials were sterilized at 160°C for 2 hours in Hot Air Oven. Spatulas were wrapped in aluminum foil and sterilized at 160°C for 2 hours in Hot Air Oven.

After sterilization, sampling vials and spatulas were kept in clean and dried sampling box (Prior cleaned with 70% Iso propyl alcohol).The sampling box were packed in plastic bag and transported to warehouse by carrying the sampling box by Sampling Personnel.

Sampling Procedure

Ø  Sampling were done under RLAF of Sampling Booth.

Ø  Sampling vials and spatulas were taken out from sampling box under RLAF after sanitizing hands with 70% IPA and wearing sterile gloves by sampling Personnel.

Ø  Sample containers were opened under RLAF.

Ø  Sampling vials were opened and sampling was done with sterile spatula and vials were closed carefully to prevent from Contamination.

Ø  The vials containing sample were kept in sampling box and the lid of sampling box were closed.

Ø  The sampling box were kept in Plastic Bag.

a.      Transport Method

Ø  The plastic bag containing sampling box were carried from warehouse to Microbiology Lab by Sampling Personnel.

Ø  The samples were stored at 2-8°C in refrigerator until tested.

Analysis Method

1 gm of sample in was dissolved in 10 ml sterile Soyabean casein digest medium and 0.01g polylobate 80 was added to sample containing tube. Using sterile pipette tips, 1ml sample was added in 90 mm diameter of sterile Petri dishes in Triplicate. 15-20 ml sterile and previously cooled at about 45°C Soyabean Casein Digest Agar (SCDA) was added in Petri dishes containing the sample for total aerobic microbial count and swirled to mix the sample. Positive Control was performed using Bacillus Subtilis ATCC 6633 and Negative control without microorganism with test simultaneously. The Petri dishes were allowed to solidify and SCDA Petri dishes were incubated at 35°C for 3 days.

Analytical Result

S.No.	Sample	Control No.	Remarks
			Total Aerobic Microbial Count (Cfu/g)
1	Microcrystalline Cellulose PH 101	A	10
2	Microcrystalline Cellulose PH 101	B	20
3	Microcrystalline Cellulose PH 101	C	10
Limit			NMT 1000 Cfu/g

 Conclusion

After analyzing, the Sample of three different control number ((A, B and C) of Microcrystalline Cellulose 101, the result was found to be in defined limit. The data obtained from the test indicates that the transport method employed for transport of microbial test sample after sampling is accurate and it has no any significant effect of contamination during transport of sample. Hence the transport method is verified and can be used to transport of microbial test samples.

 References

1. United States Pharmacopoeial Convention. (2025). <61> Microbiological examination of

Non-sterile products: Microbial enumeration tests. In USP–NF 2025. Rockville, MD.

2. Annexure 4, WHO guidelines for sampling of pharmaceutical products and related materials.

WHO Technical Report Series, No. 929, 2005.

3. Appendix 7, TRS 992. Non-sterile process validation .Annex 3, 2015 (3).

Microbial Mapping of Personnel in Class D Areas: A Risk-Based Approach to Glove Monitoring

 The “finger-dab test” is a critical, routine environmental assessment tool utilized in the Class D Manufacturing Environment (typically GMP Grade D or ISO-8), which assists in gauging the efficacy of personnel hygiene and cleaning methodologies. While there are no defined maximums by WHO TRS No. 961 for finger-dabs collected from Class D Manufacturing Facilities, most in-house action limits are established at 100 CFU (Colony Forming Units) from 5 fingerprints collected per operator.

1.      Objectives: 

To determine the risk of an increase in microbial load within finished products through manufacturing personnel during the formulation process.

2.      Study Design:

a. Design and Implementation: 1-day study completed during processing time in Class D Areas.

b. Media used: Soybean casein digest agar

c. Incubation temperature: 35°C.

d. Incubation time: 72 hours.

All samples must be collected from the Classified Areas (Class D)

 3.. Criteria for Sampling:

 1. Total samples in processing areas

2. Samples taken from right and left hands of 10 operators.

  4. Sites of Sample Collection:

 a. Granulation Room

 b. Compression Room

 c. Suspension Preparation Room

 d. Coating Room e.

Tablet Inspection Room

      5. Methods of Sample Selection

Random sampling method.

 6. Sample Collection & Analysis 

The sterile culture medium, Soybean Casein Digest Agar, was opened in the specified area; the operators were asked to gently place all five fingers of their right and left hands, respectively, in two different petri dishes. The dishes were then labeled "Right hand," "Left hand," Employee Code, and Date. A total of 20 Soybean Casein Digest Agar plates were sampled and transported to the Microbiology laboratory for analysis. Samples were incubated at 35°C for 72 hours, after which results were recorded.

6.Result

S. No	Code No.	TAMC(Cfu /Plate)	TAMC(Cfu /Plate)
		Right Hand	Left Hand
1	A	13	70
2	B	89	87
3	C	34	28
4	D	40	56
5	E	48	58
6	F	49	49
7	G	86	101
8	H	98	97
9	I	37	88
10	J	67	93

The operators succeeded in the finger test with <100 CFU on both the right and left hands when performing formulation activities in Class D work areas. However, the results are approaching the alarming limit for Class D areas of WHO TRS No. 961 Environmental Monitoring Limits. A microbial limit test is applied to non-sterile products, and the hygiene of the operators' hands plays an important role in preventing product contamination. The data demonstrate that the probability of the operator transferring their contaminating load onto the glove is high, resulting in increased microbial contamination of the end product and raising potential concerns for consumer safety.

8. Conclusion

1. Operators with high microbial loads on gloves put products at risk by adding microbes to finished products.

2. Proper methods to sanitize gloves among operators are essential to meeting the requirements for safety and quality products.

9. Recommendation

During the next revision of the regulatory guidelines for Environmental Monitoring, regulatory organizations globally, including the WHO, should make the Finger Dab Test mandatory for formulation areas in Class D and specify allowable limits for the test in the revised guidelines.

 

 

Risk-Based Module for Selecting Air Monitoring Sites for Environmental Monitoring Programme

Environmental monitoring is crucial. It’s how we keep tabs on both live microbes and non-viable particles floating in controlled, classified spaces within pharmaceutical facilities. “Controlled and classified” refers to areas defined by WHO standards, where different cleanliness levels are required for different activities. WHO divides clean rooms into Class A, B, C, and D. We can find these classifications in Annex 6 of the WHO Good Manufacturing Practice for pharmaceuticals (Technical Report No. 961, 2011).

 Why use a risk-based approach to pick sampling locations?

Because environmental monitoring isn’t just a formality. Both USP-NF (1116) and WHO guidelines stress the importance of choosing sampling sites carefully—don’t just place plates anywhere and call it monitoring. Many pharmaceutical companies overlook this and end up sampling the wrong areas, missing the true picture of microbial loads in their environments. Every facility is unique. Air exchange rates, HEPA filter performance, facility layout, activities conducted, and even operator hygiene—all influence what you find and where you find it.

 Here’s how to assemble a risk study team:

-  Members from Quality Control.

- Representatives from Quality Assurance.

- Representative from Production.

 Select your study locations thoughtfully. Focus on areas where contamination risk is higher or cleanliness is especially important. Typically, this includes:

- Dispensing room

- Granulation room

- Compression room

- Suspension preparation room

- Tablet inspection room

- Primary packing room

 Inside each room, target:

- Areas near operating machines

- High-traffic spots

- Difficult-to-clean corners

- Locations near return air vents

- Spots near drains—common sources of contamination

- Any other problem areas you observe

 How to conduct the test:

Test: Settle Plate Method

Test Condition: Dynamic

Duration: At least 3 days

Media: Soybean Casein Digest Agar (for total aerobic microbes) and Sabouraud Dextrose Agar (for yeast and mold)

Exposure time: 4 hours per plate

Incubation: Soybean Casein Digest Agar—48 to 72 hours; Sabouraud Dextrose Agar—5 to 7 days

 Steps:

1.      Place plates of each medium at your chosen sites using a petriplate stand.

                                                          Fig: Culture plate on Stand

2. Leave them open for 4 hours.

3. Carefully retrieve the plates and transport them to the microbiology lab, maintaining sterility.

4. Incubate the Soybean Casein Digest Agar plates for up to 72 hours and the Sabouraud Dextrose Agar plates for up to 7 days.

5. After incubation, count the colonies with a colony counte

6. Interptetate the results

Fig: Culture media 

 Conclusion:

Review the data. Now you’ll know which spots have higher microbial counts. Use this information to finalize your sampling sites for ongoing environmental monitoring in your facility. This way, you’re not just guessing—you’re tracking what truly matters.

Water Activity: A Faster Alternative to Traditional Testing of Non-Sterile Products in the Pharmaceutical Industry

Fig: Water Activity Meter

The microbial limit test (also known as the traditional testing method) is a time-consuming test adopted by most of the pharmaceutical industry for the release of non-sterile commercial products to market after completing their manufacturing processes. The test is generally done as per USP <61> (Microbial Enumeration Tests) for total aerobic microbial count (TAMC) and total yeast/mold count (TYMC). USP <62> (Tests for Specified Microorganisms) for the presence or absence of specific objectionable pathogens. Total time consumed to complete the test is between 5 and 7 days. The cost of the test is high because of consumption time, manpower, culture media, and the utilization of different equipment for a single test.

Water activity (aw) is the ratio of vapor pressure of H₂O in product (P) to vapor pressure of pure H₂O (Po) at the same temperature. It is numerically equal to 1/100 of the relative humidity (RH). Water activity (aw) is an important parameter for the growth of microorganisms. Practically, the water activity test can be performed and completed between 1- 2 hours. So we can say the test is less time-consuming and also a cost-effective test in comparison to the microbial limit test. Also single equipment is used for the test.

Rationale for selection of water activity (aw) as an alternate method

Most of the bacteria isolated from compressed tablets grow above water activity (aw 0.75), and yeast and molds above (aw 0.60). (Ref. USP. 35, (1112) “Application to water activity determination of non-sterile pharmaceutical products”) Estimated water activity (aw) of compressed tablets is between 0.30 and 0.40, which is very low for vegetative microorganisms to grow and multiply, but the spore-forming organism, if present in compressed tablets, can remain in a dormant phase, which can grow as a colony during the microbial limit test due to getting a favorable environment.

Assessment study of compressed tablets for water activity test and comparison with microbial limit test.

Microbial tests were done as per USP <61> (Microbial Enumeration Tests) and USP <62> (Tests for Specified Microorganisms). A water activity test of compressed tablets was done by crushing the compressed tablets in a clean, dry mortar and pestle, and the test was done using a water activity meter (Novasina). Before conducting the water activity test, the equipment was calibrated using SAL-T 58, SAL-T 11, and SAL-T 84  standard provided by the manufacturer.

Result

S.No	Products	Observed Value (Initial test)		Observed Value (12 month Real time stability)
		Water activity (aw)	Microbial Limit Test	Water activity (aw)	Microbial Limit Test
1	Chlorthalidone 6.25 mg tablet	0.430@25.8°C	10 Cfu/g	0.433@25.1°C	10 Cfu/g
2	Chlorthalidone 12.5 mg tablet	0.315@25.4°C	0 Cfu/g	0.327@24.9°C	10 Cfu/g
3	Chlorthalidone 25 mg tablet	0.346@25.9°C	20 Cfu/g	0.356@25.0°C	20 Cfu/g
4	Amlodipine 5 mg tablet	0.511@25.6°C	30 Cfu/g	0.510@25.2°C	20 Cfu/g
5	Methylcobalamin 1500 mcg tablet	0.432@25.2°C	10 Cfu/g	0.427@25.1°C	10 Cfu/g

Result Interpretation 

Small sample size studies were done under laboratory conditions for comparison between the microbial limit test and water activity (aw) to find the relation of water activity for microbial growth. The result was found in agreement with USP 35, (1112) “Application to water activity determination of non-sterile pharmaceutical products,” as the growth of organisms did not significantly increase in the test result. The water activity was found below 0.75 in all samples, and the load of organisms during the initial test and 12-month real-time stability did not vary drastically.

Conclusion 

Based on our test result, we can conclude that the water activity test can be used as an alternate method for testing compressed tablets in the pharmaceutical industry for immediate batch release, changing the testing method to an automated method and saving time and cost to increase productivity and cost-effectiveness within the pharmaceutical industry. 

Limitation of study 

1. The sample size of the study is too small to draw any realistic conclusion.

 Recommendation

 Any manufacturing industry that wants to implement water activity as an alternate method  to the microbial limit test ,is suggested to test the large sample size under their own laboratory condition before implementation of alternate method.